Process of making somatostatin modulators

ABSTRACT

Described herein are compounds that are somatostatin modulators, methods of making such compounds, pharmaceutical compositions and medicaments comprising such compounds, and methods of using such compounds in the treatment of conditions, diseases, or disorders that would benefit from modulation of somatostatin activity.

RELATED APPLICATIONS

This application is a continuation application of U.S. application Ser.No. 16/249,729, filed Jan. 16, 2019, which claims the benefit of U.S.Provisional Patent Application No. 62/618,538 filed on Jan. 17, 2018,each of which is herein incorporated by reference in its entirety.

FIELD OF THE INVENTION

Described herein are compounds that are somatostatin modulators, methodsof making such compounds, pharmaceutical compositions and medicamentscomprising such compounds, and methods of using such compounds in thetreatment of conditions, diseases, or disorders that would benefit frommodulating somatostatin activity.

BACKGROUND OF THE INVENTION

Somatostatin is a peptide hormone that regulates the endocrine systemand affects neurotransmission and cell proliferation via interactionwith G-protein-coupled somatostatin receptors and inhibition of therelease of numerous secondary hormones. Six subtype somatostatinreceptor proteins have been identified (SSTR1, SSTR2a, SSTR2b, SSTR3,SSTR4, SSTR5) and are encoded by five different somatostatin receptorgenes. Modulation of a particular subtype somatostatin receptor, orcombination thereof, is attractive for the treatment of conditions,diseases, or disorders that would benefit from modulating somatostatinactivity.

SUMMARY OF THE INVENTION

In one aspect, described herein the compound3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrilemonohydrochloride, or solvate thereof, having the following structure:

In another aspect, described herein is crystalline3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrilemonohydrochloride, or solvate thereof. In some embodiments, thecrystalline3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrilemonohydrochloride, or solvate thereof, has: an X-ray powder diffraction(XRPD) pattern with peaks at 4.5° 2-Theta, 9.1° 2-Theta, 10.2° 2-Theta,16.3° 2-Theta, 18.4° 2-Theta, and 19.1° 2-Theta; an X-ray powderdiffraction (XRPD) pattern substantially the same as shown in FIG. 1; aDifferential Scanning Calorimetry (DSC) thermogram with an endothermhaving an onset at about 207° C. and a peak at about 220° C.; aDifferential Scanning Calorimetry (DSC) thermogram substantially thesame as shown in FIG. 2(a); a Thermogravimetric Analysis (TGA)thermogram substantially the same as shown in FIG. 2(b); an infrared(IR) spectrum with peaks at 2223 cm⁻¹, 1620 cm⁻¹, 1595 cm⁻¹, 1457 cm⁻¹,1238 cm⁻¹, 1220 cm⁻¹, and 1117 cm⁻¹; an infrared (IR) spectrumsubstantially the same as shown in FIG. 3; an unchanged XRPD when heatedup to about 200° C., upon exposure to more than 90% relative humidityfor about 24 hours, or upon exposure to about 75% RH and 40° C. over oneweek, or combinations thereof; or combinations thereof.

In some embodiments, the crystalline3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrilemonohydrochloride, or solvate thereof, has an X-ray powder diffraction(XRPD) pattern with peaks at 4.5° 2-Theta, 9.1° 2-Theta, 10.2° 2-Theta,16.3° 2-Theta, 18.4° 2-Theta, and 19.1° 2-Theta. In some embodiments,the crystalline3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrilemonohydrochloride, or solvate thereof, has an XRPD pattern furthercomprises peaks at 20.7° 2-Theta, 23.3° 2-Theta, 23.4° 2-Theta, 23.6°2-Theta, 27.1° 2-Theta, and 28.0° 2-Theta. In some embodiments, thecrystalline3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrilemonohydrochloride, or solvate thereof, has an X-ray powder diffraction(XRPD) pattern substantially the same as shown in FIG. 1. In someembodiments, the crystalline3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrilemonohydrochloride, or solvate thereof, has a Differential ScanningCalorimetry (DSC) thermogram with an endotherm having an onset at about207° C. and a peak at about 220° C. In some embodiments, the crystalline3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrilemonohydrochloride, or solvate thereof, has a Differential ScanningCalorimetry (DSC) thermogram substantially the same as shown in FIG.2(a). In some embodiments, the crystalline3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrilemonohydrochloride, or solvate thereof, has an unchanged XRPD when heatedup to about 200° C. In some embodiments, the crystalline3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrilemonohydrochloride, or solvate thereof, has an unchanged XRPD uponexposure to more than 90% relative humidity for 24 hours and uponexposure to about 75% RH and 40° C. over one week. In some embodiments,the crystalline3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrilemonohydrochloride, or solvate thereof, has an infrared (IR) spectrumwith characteristic peaks at 2223 cm⁻¹, 1620 cm⁻¹, 1595 cm⁻¹, 1457 cm⁻¹,1238 cm⁻¹, 1220 cm⁻¹, and 1117 cm⁻¹. In some embodiments, thecrystalline3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrilemonohydrochloride, or solvate thereof, has an infrared (IR) spectrumsubstantially the same as the IR spectrum shown in FIG. 3.

In another aspect, described herein is a pharmaceutical compositioncomprising crystalline3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrilemonohydrochloride, or solvate thereof, and at least one pharmaceuticallyacceptable excipient. In some embodiments, the pharmaceuticalcomposition is formulated for administration to a mammal by oraladministration. In some embodiments, the pharmaceutical composition isin the form of a solid form pharmaceutical composition. In someembodiments, the pharmaceutical composition is in the form of a tablet,a pill, or a capsule.

In another aspect, described herein is a method of making crystalline3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrilemonohydrochloride, or solvate thereof, comprising the steps of:

-   -   (a) slurrying        3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrile        dihydrochloride in 5 volumes of isopropanol:water (1:1) mixture;        -   (i) heating the slurry of (a) to about 45° C.;        -   (ii) adding about 0.5 to about 1.2 equivalents of ammonium            hydroxide solution, sodium bicarbonate solution, or sodium            hydroxide solution to the heated slurry of step (a)(i) to            achieve a pH of about 4.0-6.0;        -   (iii) adding water over about 2 hours to the mixture of step            (a)(ii); and        -   (iv) filtering the slurry of step (a)(iii) to provide            3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrile            monohydrochloride, or solvate thereof;    -   or    -   (b) adding a suitable solvent to        3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrile;        -   (i) adding about 1 equivalent of hydrochloric acid to the            mixture of solvent and            3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrile            of (b); and        -   (ii) filtering the solids resulting from step (b)(ii) to            provide            3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrile            monohydrochloride, or solvate thereof;    -   or    -   (c) stirring        3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrile        dihydrochloride in about 20 volumes to about 50 volumes of        water; and        -   (i) filtering the solids of step (c) to provide            3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrile            monohydrochloride, or solvate thereof.

In some embodiments, ammonium hydroxide solution is used in (a)(ii). Insome embodiments, the amount of ammonium hydroxide solution used in(a)(ii) is about 0.8 equivalents and the pH achieved is about 4.5-4.7.

In some embodiments, the suitable solvent in (b) is methanol, ethanol,isopropyl alcohol, acetone, methyl acetate, ethyl acetate,tetrahydrofuran, tetrahydropyran, water, or combinations thereof.

In another aspect, described herein is a process for the synthesis of3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitriledihydrochloride:

comprising the step of treating Compound A-VI:

with hydrochloric acid in a suitable solvent.

In another aspect, described herein is a process for the synthesis of3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrilemonohydrochloride:

comprising the steps of:

-   -   (1) treating Compound A-VI:

-   -   with hydrochloric acid in a suitable solvent to provide        3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrile        dihydrochloride; and    -   (2) treating        3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrile        dihydrochloride with aqueous ammonia to provide        3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrile        monohydrochloride.

In some embodiments, the suitable solvent is isopropyl alcohol, ethylacetate, or isopropyl acetate. In some embodiments, the suitable solventis isopropyl alcohol.

In another aspect, described herein is a process for the preparation ofCompound A-VI:

comprising the steps of:

-   -   (1) reacting Compound A-IV:

-   -   -   with Compound 1:

-   -   -   wherein,            -   B is a boronic acid, boronate ester, or trifluoroborate;        -   in the presence of a coupling catalyst, a suitable base, and            in a suitable solvent, to provide Compound A-V:

-   -   and    -   (2) reacting Compound A-V with 3,5-difluorophenylboronic acid:

-   -   in the presence of a coupling catalyst, a suitable base, and in        a suitable solvent, to provide Compound A-VI.

In some embodiments, B is a boronic acid or trifluoroborate.

In some embodiments, B is a boronic acid. In some embodiments, B istrifluoroborate.

In some embodiments, the coupling catalyst of step (1) is a palladiumcatalyst; the suitable base of step (1) is triethylamine,diisopropylethylamine, 1,2,2,6,6-pentamethylpiperidine, tributylamine,sodium bicarbonate, Na₂CO₃, K₂CO₃, Cs₂CO₃, NaOAc, KOAc, Ba(OH)₂, Na₃PO₄or K₃PO₄; and the suitable solvent of step (1) is acetonitrile,dimethylformamide, ethanol, tetrahydrofuran, isopropyl alcohol,1,4-dioxane, water, or combinations thereof. In some embodiments, Step(1) is performed at a temperature of about 80° C. In some embodiments,Step (1) is performed at a temperature of about 80-85° C.

In some embodiments, the coupling catalyst of step (1) is a palladiumcatalyst; the suitable base of step (1) is K₂CO₃; and the suitablesolvent of step (1) is a mixture of 1,4-dioxane and water.

In some embodiments, the coupling catalyst of step (2) is a palladiumcatalyst; the suitable base of step (2) is triethylamine,diisopropylethylamine, 1,2,2,6,6-pentamethylpiperidine, tributylamine,sodium bicarbonate, Na₂CO₃, K₂CO₃, Cs₂CO₃, NaOAc, KOAc, Ba(OH)₂, Na₃PO₄or K₃PO₄; and the suitable solvent of step (2) is acetonitrile,dimethylformamide, ethanol, tetrahydrofuran, isopropyl alcohol,1,4-dioxane, water, or combinations thereof. In some embodiments, Step(2) is performed at a temperature of about 90° C. to about 100° C. Insome embodiments, the coupling catalyst of step (2) is a palladiumcatalyst; the suitable base of step (2) is K₂CO₃; and the suitablesolvent of step (2) is a mixture of 1,4-dioxane and water.

In some embodiments, Compound A-V is isolated prior to step (2).

In some embodiments, Compound A-V is not isolated prior to step (2).

In some embodiments, the process further comprises recrystallizingCompound A-VI from a suitable solvent. In some embodiments, the suitablesolvent is methyl acetate, ethyl acetate, isopropyl acetate, methanol,ethanol, isopropyl alcohol, dichloromethane/petroleum ether,acetonitrile, tetrahydrofuran/water, tetrahydrofuran/petroleum ether,dimethylformamide/water, dichloromethane/methyl tert-butyl ether,methanol/methyl tert-butyl ether, methyl tert-butyl ether, or toluene.In some embodiments, the suitable solvent is ethyl acetate or isopropylacetate.

In some embodiments, the process further comprises treatment of therecrystallized Compound A-VI with a metal scavenger. In someembodiments, the metal scavenger comprises SiO₂, charcoal, aqueoussolution of L-cysteine, a Silicycle metal scavenger, Si-thiol, SiliaBondDMT or SiliaBond Cysteine.

In another aspect, described herein is a process for the preparation ofCompound A-IV:

comprising the steps of:

-   -   (1) chlorinating Compound A-I:

-   -   with a suitable chlorinating agent in a suitable solvent to        provide Compound A-II:

-   -   (2) brominating Compound A-II with a suitable brominating agent        in a suitable solvent to provide Compound A-III:

-   -   and    -   (3) coupling 4-(N-Boc amino)piperidine with Compound A-III in        the presence of a suitable base and in a suitable solvent to        provide Compound A-IV;    -   or    -   (i) coupling 4-(N-Boc amino)piperidine with        6-bromo-4-chloro-quinoline in the presence of a suitable base        and in a suitable solvent to provide Compound 4:

-   -   and    -   (ii) chlorinating Compound 4 with a suitable chlorinating agent        in a suitable solvent to provide Compound A-IV.

In some embodiments, the chlorinating agent of step (1) isN-chlorosuccinimide, trichloroisocyanuric acid, sulfuryl chloride,chlorine, sodium hypochlorite, calcium hypochlorite, hypochlorous acid,or 2,3,4,5,6,6-hexachloro-2,4-cyclohexadien-1-one; and the suitablesolvent of step (1) is acetic acid, water, ethanol, methanol, toluene,dichloromethane, tetrahydrofuran, dioxane, or N,N-dimethylformamide.

In some embodiments, the chlorinating agent of step (1) isN-chlorosuccinimide; and the suitable solvent of step (1) is aceticacid.

In some embodiments, the brominating agent of step (2) is phosphorustribromide, phosphorus oxybromide, hydrobromic acid, bromine, ordibromotriphenylphosphorane; and the suitable solvent of step (2) isacetonitrile, water, ethanol, isopropanol, dichloromethane, toluene,N,N-dimethylformamide, acetic acid, or acetone.

In some embodiments, the brominating agent of step (2) is phosphorustribromide; and the suitable solvent of step (2) isN,N-dimethylformamide.

In some embodiments, the suitable base of step (3) is triethylamine,diisopropylethylamine, 1,8-diazabicycloundec-7-ene,1,2,2,6,6-pentamethylpiperidine, tributylamine, sodium bicarbonate,Na₂CO₃, K₂CO₃, or Cs₂CO₃; and the suitable solvent of step (3) isN,N-dimethylformamide, N,N-dimethylacetamide, dimethylsulfoxide,dichlormethane, chloroform, carbon tetrachloride, dioxane,tetrahydrofuran, toluene, acetonitrile, ethanol, or isopropanol.

In some embodiments, the base of step (3) is diisopropylethylamine; andthe suitable solvent of step (3) is dimethylsulfoxide.

In some embodiments, the suitable base of step (i) is triethylamine,diisopropylethylamine, 1,8-diazabicycloundec-7-ene,1,2,2,6,6-pentamethylpiperidine, tributylamine, sodium bicarbonate,Na₂CO₃, K₂CO₃, or Cs₂CO₃; and the suitable solvent of step (i) isN,N-dimethylformamide, N,N-dimethylacetamide, dimethylsulfoxide,dichlormethane, chloroform, carbon tetrachloride, dioxane,tetrahydrofuran, toluene, acetonitrile, ethanol, or isopropanol.

In some embodiments, the base of step (i) is K₂CO₃; and the suitablesolvent of step (i) is N,N-dimethylformamide.

In some embodiments, the chlorinating agent of step (ii) isN-chlorosuccinimide, trichloroisocyanuric acid, sulfuryl chloride,chlorine, sodium hypochlorite, calcium hypochlorite, hypochlorous acid,or 2,3,4,5,6,6-hexachloro-2,4-cyclohexadien-1-one; and the suitablesolvent of step (ii) is acetic acid, water, ethanol, methanol, toluene,dichloromethane, tetrahydrofuran, dioxane, or N,N-dimethylformamide.

In some embodiments, the chlorinating agent of step (ii) isN-chlorosuccinimide; and the suitable solvent of step (ii) is toluene.

In another aspect, described herein is a process for the preparation ofCompound 1:

wherein,

-   -   B is a boronic acid or boronate ester;

comprising the steps of:

(1) protecting the hydroxyl group of Compound 2:

with a suitable protecting group (PG′) to provide Compound 2a:

(2) reacting Compound 2a with a borylation agent under suitable reactionconditions; and

(3) removal of the protecting group (PG′) to provide Compound 1.

In some embodiments, B is a boronic acid.

In some embodiments, the process further comprises converting B to atrifluoroborate.

In some embodiments, the borylation agent is triisopropyl borate,trimethyl borate, tetrahydroxydiboron, pinacolborane, catecholborane,bis(neopentyl glycolato)diboron, bis(pinacolato)diboron, bis(hexyleneglycolato)diboron, bis(catecholato)diboron,4,4,5,5-tetramethyl-1,3,2-dioxaborolane,4,6,6-trimethyl-1,3,2-dioxaborinane, diisopropylamine borane,bis(neopentyl glycolato)diboron, bis(catecholato)diboron, orbis(pinacolato)diboron.

In some embodiments, the suitable reaction conditions of (2) comprisethe use of metal halogen exchange reagents. In some embodiments, thesuitable reaction conditions of (2) comprise the use of metal halogenexchange reagents selected from Grignard reagents and alkyl lithiumreagents. In some embodiments, the suitable reaction conditions of (2)comprise the use of isopropyl magnesium chloride in tetrahydrofuran.

In some embodiments, the borylation agent is triisopropyl borate and thesuitable reaction conditions of (2) comprise the use of isopropylmagnesium chloride in tetrahydrofuran.

In some embodiments, the suitable reaction conditions of (2) comprisethe use of transition metal mediated reaction conditions.

In some embodiments, the suitable reaction conditions of (2) comprisethe use of palladium metal mediated reaction conditions.

In some embodiments, the suitable protecting group (PG′) ismethoxymethyl, ethoxyethyl, methoxypropyl, benzyloxymethyl,2-methoxyethoxymethyl, benzyl, para-methoxybenzyl, 2-naphthylmethyl,methyl, allyl, tetrahydropyranyl, acetyl, benzoyl, 2,2,2-trichloroethylcarbonyl, trimethylsilyl, triethylsilyl, triisopropyl silyl,tert-butyldimethylsilyl, or tert-butyldiphenylsilyl.

In some embodiments, the suitable protecting group (PG′) ismethoxymethyl, 2-methoxyethoxymethyl, benzyl, para-methoxybenzyl,methyl, allyl, tetrahydropyran-2-yl, [2-(trimethylsilyl)ethoxy]methyl,trimethylsilyl, triethylsilyl, triisopropyl silyl,tert-butyldimethylsilyl, or tert-butyldiphenylsilyl.

In some embodiments, the suitable protecting group (PG′) ismethoxymethyl, ethoxyethyl, methoxypropyl, benzyloxymethyl,2-methoxyethoxymethyl, benzyl, para-methoxybenzyl, 2-naphthylmethyl,methyl, allyl, or tetrahydropyranyl. In some embodiments, the suitableprotecting group (PG′) is methoxymethyl, ethoxyethyl, or2-methoxyethoxymethyl. In some embodiments, the suitable protectinggroup (PG′) is methoxymethyl.

In some embodiments, the suitable protecting group (PG′) is acetyl,benzoyl, or 2,2,2-trichloroethyl carbonyl. In some embodiments, thesuitable protecting group (PG′) is acetyl.

In some embodiments, removal of the protecting group in step (3) isaccomplished by treatment with hydrochloric acid, hydrobromic acid,acetic acid, trifluoroacetic acid, para-toluenesulfonic acid, ZnBr₂,hydrogen over Pd/C, 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ),boron tribromide, boron trichloride, trimethylsilyl iodide, Pd(PPh₃)₄,tetra-n-butylammonium fluoride (TBAF), or HF-pyridine.

In some embodiments, the suitable protecting group (PG′) ismethoxymethyl; and removal of the protecting group in step (3) isaccomplished by treatment with hydrochloric acid.

Articles of manufacture, which include packaging material, asomatostatin modulator, or a pharmaceutically acceptable salt or solvatethereof, as described herein, within the packaging material, and a labelthat indicates that the somatostatin modulator, or pharmaceuticallyacceptable salt, or solvate thereof, is used for modulating one or moresubtype somatostatin receptor proteins, or for the treatment, preventionor amelioration of one or more symptoms of a disease or condition thatwould benefit from modulating one or more subtype somatostatin receptorproteins, are provided.

Other objects, features and advantages of the compounds, methods andcompositions described herein will become apparent from the followingdetailed description. It should be understood, however, that thedetailed description and the specific examples, while indicatingspecific embodiments, are given by way of illustration only, sincevarious changes and modifications within the spirit and scope of theinstant disclosure will become apparent to those skilled in the art fromthis detailed description.

BRIEF DESCRIPTION OF THE DRAWINGS

FIG. 1. X-ray powder diffraction (XRPD) pattern of Compound A, mono-HClsalt.

FIG. 2(a). Differential Scanning Calorimetry (DSC) thermogram.

FIG. 2(b). Thermogravimetric Analysis/Differential Scanning Calorimetry(TGA/DSC) thermogram of Compound A, mono-HCl salt.

FIG. 3. Infrared (IR) spectrum of Compound A, mono-HCl salt.

FIG. 4. X-ray powder diffraction (XRPD) pattern of Compound A, di-HClsalt.

FIG. 5(a). Differential Scanning Calorimetry (DSC) thermogram ofCompound A, di-HCl salt.

FIG. 5(b). Thermogravimetric Analysis (TGA) thermogram of Compound A,di-HCl salt.

FIG. 6. Infrared (IR) spectrum of Compound A, di-HCl salt.

FIG. 7(a). X-ray powder diffraction (XRPD) pattern A of Compound A, freebase.

FIG. 7(b). X-ray powder diffraction (XRPD) pattern B of Compound A, freebase.

FIG. 7(c). X-ray powder diffraction (XRPD) pattern C of Compound A, freebase.

FIG. 8(a). Differential Scanning Calorimetry (DSC) thermogram of PatternC of Compound A, free base.

FIG. 8(b). Thermogravimetric Analysis (TGA) thermogram of Pattern C ofCompound A, free base.

FIG. 9. X-ray powder diffraction (XRPD) pattern of Compound A, mono-HClsalt before (bottom spectra) and after (top spectra) Dynamic VaporSorption (DVS) testing between 2 and 95% Relative Humidity (RH).

FIG. 10. X-ray powder diffraction (XRPD) pattern of Compound A, di-HClsalt before (bottom spectra) and after (top spectra) Dynamic VaporSorption (DVS) testing between 2 and 95% Relative Humidity (RH).

DETAILED DESCRIPTION OF THE INVENTION

Somatostatin (SST), also known as somatotropin release inhibiting factor(SRIF) was initially isolated as a 14-amino acid peptide from ovinehypothalamii (Brazeau et al., Science 179, 77-79, 1973). An N-terminalextended 28-amino acid peptide with similar biological activity to14-amino acid somatostatin was subsequently isolated (Pradayrol et, al.,FEBS Letters, 109, 55-58, 1980; Esch et al., Proc. Natl. Acad. Sci. USA,77, 6827-6831, 1980). SST is a regulatory peptide produced by severalcell types in response to other neuropeptides, neurotransmitters,hormones, cytokines, and growth factors. SST acts through both endocrineand paracrine pathways to affect its target cells. Many of these effectsare related to the inhibition of secretion of other hormones, mostnotably growth hormone (GH). They are produced by a wide variety of celltypes in the central nervous system (CNS) and gut, and have multiplefunctions including modulation of secretion of growth hormone (GH),insulin, glucagon, as well as many other hormones that areanti-proliferative.

These pleotropic actions of somatostatins are mediated by sixsomatostatin receptor proteins (SSTR1, SSTR2a, SSTR2b, SSTR3, SSTR4,SSTR5). The six somatostatin receptor proteins are encoded by fivedifferent somatostatin receptor genes (Reisine and Bell, Endocr Rev. 16,427-442, 1995; Patel and Srikant, Trends EndocrinolMetab 8, 398-405,1997). All the receptors are members of the class-A subgroup of the GPCRsuperfamily. SST2A receptor is the most widely expressed subtype inhuman tumors and is the dominant receptor by which GH secretion issuppressed. Unless otherwise stated, the term SSTR2 means SSTR2a.

It is possible to selectively modulate any one of the somatostatinreceptor subtypes, or combination thereof. In some embodiments,selectively modulating any one of the somatostatin receptor subtypesrelative to the other somatostatin receptor subtypes, or combinationthereof, is useful in a variety of clinical applications. In someembodiments, selectively modulating any one of the somatostatin receptorsubtypes relative to the other somatostatin receptor subtypes reducesunwanted side effects in a variety of clinical applications.

For example, modulation of SSTR2 activity mediates the inhibition ofgrowth hormone (GH) release from the anterior pituitary and glucagonrelease from pancreas. SSTR2 is also implicated in many other biologicalfunctions such as, but not limited to, cell proliferation, nociception,inflammation, and angiogenesis. In some embodiments, a selective SSTR2modulator is used in the treatment of acromegaly, gut neuroendocrinetumors, pain, neuropathies, nephropathies, and inflammation, as well asretinopathies resulting from aberrant blood vessel growth. In some otherembodiments, a selective SSTR2 modulator is used in the treatment ofarthritis, pain, cancer, inflammatory bowel disease, irritable bowelsyndrome, Crohn's disease, Cushing's disease, acute lung injury, acuterespiratory distress syndrome, and ophthalmic disorders such asage-related macular degeneration (AMD), diabetic retinopathy, diabeticmacular edema, and Graves ophthalmology, among others.

In some embodiments, SSTR4 agonists exhibit anti-inflammatory andanti-nociceptive effects.

In some embodiments, SSTR3 agonists inhibit insulin secretion.

In some embodiments, SSTR5 agonists inhibit insulin secretion. Inaddition, SSTR5 has also been implicated to modulate the release ofgrowth hormone.

Somatostatin peptide and its receptor subtypes are also widely expressedin the brain and disruption or diminishment of their activity ispotentially involved in several psychiatric and neurodegenerativediseases. For example, concentrations of somatostatin in the cerebralcortex and hippocampus are reduced in schizophrenics and one of the mostconsistent neuropathologic findings in this patient group is a deficitin cortical inhibitory interneurons expressing somatostatin.Somatostatin is also highly expressed in brain regions associated withseizures and has also been implicated as having an important role inepilepsy. Somatostatin levels are diminished in the hippocampi ofAlzheimer's and Parkinson's patients, suggesting that restoration of itssignaling as a potential drug target for neurodegeneration.

In one aspect, compounds described herein are modulators of SSTR2. Insome embodiments, compounds described herein selectively modulate theactivity of SSTR2 relative to the other somatostatin receptors.

In some embodiments, compounds described here are amenable to oraladministration to a mammal in need of treatment with a somatostatinmodulator.

In some embodiments, somatostatin receptor modulators described hereinhave utility over a wide range of therapeutic applications. In someembodiments, somatostatin receptor modulators described herein are usedin the treatment of a variety of diseases or conditions such as, but notlimited to acromegaly, neuroendocrine tumors, retinopathies and otherophthalmic disorders, neuropathy, nephropathy, respiratory diseases,cancers, pain, neurodegenerative diseases, inflammatory diseases, aswell as psychiatric and neurodegenerative disorders. In someembodiments, somatostatin receptor modulators described herein are usedin the treatment of acromegaly in a mammal.

In some embodiments, somatostatin receptor modulators described hereininhibit the secretion of various hormones and trophic factors inmammals. In some embodiments, the compounds are used to suppress certainendocrine secretions, such as, but not limited to GH, insulin, glucagonand prolactin. The suppression of certain endocrine secretions is usefulin the treatment of disorders such as acromegaly; endocrine tumors suchas carcinoids, VIPomas, insulinomas and glucagonomas; or diabetes anddiabetes-related pathologies, including retinopathy, neuropathy andnephropathy. In some embodiments, somatostatin receptor modulatorsdescribed herein are used to suppress exocrine secretions in thepancreas, stomach and intestines, for the treatment of disorders such aspancreatitis, fistulas, bleeding ulcers and diarrhea associated withsuch diseases as AIDS or cholera. Disorders involving autocrine orparacrine secretions of trophic factors such as IGF-1 (as well as someendocrine factors) which may be treated by administration of thecompounds described herein include cancers of the breast, prostate, andlung (both small cell and non-small cell epidermoids), as well ashepatomas, neuroblastomas, colon and pancreatic adenocarcinomas (ductaltype), chondrosarcomas, and melanomas, diabetic retinopathy, andatherosclerosis associated with vascular grafts and restenosis followingangioplasty.

In some embodiments, somatostatin receptor modulators described hereinare used to suppress the mediators of neurogenic inflammation (e.g.substance P or the tachykinins), and may be used in the treatment ofrheumatoid arthritis; psoriasis; topical inflammation such as isassociated with sunburn, eczema, or other sources of itching;inflammatory bowel disease; irritable bowel syndrome; allergies,including asthma and other respiratory diseases In some otherembodiments, the somatostatin receptor modulators described hereinfunction as neuromodulators in the central nervous system and are usefulin the treatment of Alzheimer's disease and other forms of dementia,pain, and headaches. In some embodiments, somatostatin receptormodulators described herein provide cytoprotection in disordersinvolving the splanchnic blood flow, including cirrhosis and oesophagalvarices.

Compound A is a somatostatin modulator that is useful in the methods oftreatment described herein.

Compound A

As used herein, Compound A refers to3-(4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl)-2-hydroxy-benzonitrile,which has the chemical structure shown below.

In some embodiments, Compound A is amorphous.

In some embodiments, Compound A is crystalline.

In some embodiments, Compound A is crystalline and has an X-Ray powderdiffraction pattern with peaks at 9.2° 2-Theta, 12.3° 2-Theta, 14.4°2-Theta, and 24.0° 2-Theta. In some embodiments, Compound A iscrystalline and has an X-Ray powder diffraction pattern substantiallysimilar to the XRPD displayed in FIG. 7(a).

In some embodiments, Compound A is crystalline and has an X-Ray powderdiffraction pattern with peaks at 5.9° 2-Theta, 13.9° 2-Theta, 14.2°2-Theta, 17.5° 2-Theta, and 24.6° 2-Theta. In some embodiments, CompoundA is crystalline and has an X-Ray powder diffraction patternsubstantially similar to the XRPD displayed in FIG. 7(b).

In some embodiments, Compound A is crystalline and is characterized ashaving: an X-ray powder diffraction (XRPD) pattern with peaks at 7.2°2-Theta, 8.3° 2-Theta, 10.9° 2-Theta, and 12.0° 2-Theta; an X-ray powderdiffraction (XRPD) pattern substantially the same as shown in FIG. 7(c);a Differential Scanning Calorimetry (DSC) thermogram with an endothermhaving an onset at about 128° C. and a peak at about 145° C.; aDifferential Scanning Calorimetry (DSC) thermogram substantially thesame as shown in FIG. 8(a); a Differential Scanning Calorimetry (DSC)thermogram substantially the same as shown in FIG. 8(a); aThermogravimetric Analysis (TGA) thermogram substantially the same asshown in FIG. 8(b); or combinations thereof.

In some embodiments, Compound A is crystalline and is characterized ashaving at least one of the following properties:

-   -   a) an X-ray powder diffraction (XRPD) pattern with peaks at 7.2°        2-Theta, 8.3° 2-Theta, 10.9° 2-Theta, and 12.0° 2-Theta;    -   b) an X-ray powder diffraction (XRPD) pattern substantially the        same as shown in FIG. 7(c);    -   c) a Differential Scanning Calorimetry (DSC) thermogram with an        endotherm having an onset at about 128° C. and a peak at about        145° C.;    -   d) a Differential Scanning Calorimetry (DSC) thermogram        substantially the same as shown in FIG. 8(a).

In some embodiments, Compound A is crystalline and is characterized ashaving at least two of the properties selected from a) to d). In someembodiments, Compound A is crystalline and is characterized as having atleast three of the properties selected from a) to d). In someembodiments, Compound A is crystalline and is characterized as havingproperties a), b), c), and d).

In some embodiments, Compound A is crystalline and has an X-ray powderdiffraction (XRPD) pattern with peaks at 7.2° 2-Theta, 8.3° 2-Theta,10.9° 2-Theta, and 12.0° 2-Theta. In some embodiments, Compound A iscrystalline and has an X-ray powder diffraction (XRPD) patternsubstantially the same as shown in FIG. 7(c). In some embodiments,Compound A is crystalline and has a Differential Scanning Calorimetry(DSC) thermogram with an endotherm having an onset at about 128° C. anda peak at about 145° C. In some embodiments, Compound A is crystallineand has a Differential Scanning Calorimetry (DSC) thermogramsubstantially the same as shown in FIG. 8(a).

In some embodiments, provided herein is a pharmaceutically acceptablesalt of Compound A. In some embodiments, the pharmaceutically acceptablesalt of Compound A is the monohydrochloride salt (Compound A-HCl):

In some embodiments, Compound A-HCl is amorphous.

In some embodiments, Compound A-HCl is crystalline.

In some embodiments, Compound A-HCl is crystalline and is characterizedas having: an X-ray powder diffraction (XRPD) pattern with peaks at 4.5°2-Theta, 9.1° 2-Theta, 10.2° 2-Theta, 16.3° 2-Theta, 18.4° 2-Theta, and19.1° 2-Theta; an X-ray powder diffraction (XRPD) pattern substantiallythe same as shown in FIG. 1; a Differential Scanning Calorimetry (DSC)thermogram with an endotherm having an onset at about 207° C. and a peakat about 220° C.; a Differential Scanning Calorimetry (DSC) thermogramsubstantially the same as shown in FIG. 2(a); a ThermogravimetricAnalysis (TGA) thermogram substantially the same as shown in FIG. 2(b);an infrared (IR) spectrum with peaks at 2223 cm⁻¹, 1620 cm⁻¹, 1595 cm⁻¹,1457 cm⁻¹, 1238 cm⁻¹, 1220 cm⁻¹, and 1117 cm⁻¹; an infrared (IR)spectrum substantially the same as shown in FIG. 3; an unchanged XRPDwhen heated up to about 200° C., upon exposure to more than 90% relativehumidity for about 24 hours, or upon exposure to about 75% RH and 40° C.over one week, or combinations thereof, or combinations thereof.

In some embodiments, Compound A-HCl is crystalline and is characterizedas having at least one of the following properties:

-   -   a) an X-ray powder diffraction (XRPD) pattern with peaks at 4.5°        2-Theta, 9.1° 2-Theta, 10.2° 2-Theta, 16.3° 2-Theta, 18.4°        2-Theta, and 19.1° 2-Theta;    -   b) an X-ray powder diffraction (XRPD) pattern substantially the        same as shown in FIG. 1;    -   c) a Differential Scanning Calorimetry (DSC) thermogram with an        endotherm having an onset at about 207° C. and a peak at about        220° C.;    -   d) a Differential Scanning Calorimetry (DSC) thermogram        substantially the same as shown in FIG. 2(a);    -   e) an infrared (IR) spectrum with characteristic peaks at 2223        cm⁻¹, 1620 cm⁻¹, 1595 cm⁻¹, 1457 cm⁻¹, 1238 cm⁻¹, 1220 cm⁻¹, and        1117 cm⁻¹;    -   f) an infrared (IR) spectrum substantially the same as shown in        FIG. 3;    -   g) reversible water uptake (˜4.5% w/w) between 2 and 95%        Relative Humidity (RH);    -   h) reversible water uptake (˜2.3%) between 15 and 75% Relative        Humidity (RH);    -   i) an unchanged XRPD after DVS analysis at 90% RH and room        temperature over 24 hours; or    -   j) an unchanged XRPD after the DVS analysis at 75% RH and 40° C.        over one week.

In some embodiments, Compound A-HCl is crystalline and is characterizedas having at least two of the properties selected from a) to j). In someembodiments, Compound A-HCl is crystalline and is characterized ashaving at least three of the properties selected from a) to j). In someembodiments, Compound A-HCl is crystalline and is characterized ashaving at least four of the properties selected from a) to j). In someembodiments, Compound A-HCl is crystalline and is characterized ashaving at least five of the properties selected from a) to j). In someembodiments, Compound A-HCl is crystalline and is characterized ashaving at least six of the properties selected from a) to j). In someembodiments, Compound A-HCl is crystalline and is characterized ashaving at least seven of the properties selected from a) to j). In someembodiments, Compound A-HCl is crystalline and is characterized ashaving at least eight of the properties selected from a) to j). In someembodiments, Compound A-HCl is crystalline and is characterized ashaving at least nine of the properties selected from a) to j). In someembodiments, Compound A-HCl is crystalline and is characterized ashaving properties a), b), c), d), e), f), g), h), i), and j).

In some embodiments, Compound A-HCl is crystalline and has an X-raypowder diffraction (XRPD) pattern with peaks at 4.5° 2-Theta, 9.1°2-Theta, 10.2° 2-Theta, 16.3° 2-Theta, 18.4° 2-Theta, and 19.1° 2-Theta.In some embodiments, Compound A-HCl is crystalline and has an X-raypowder diffraction (XRPD) pattern substantially the same as shown inFIG. 1. In some embodiments, Compound A-HCl is crystalline and has aDifferential Scanning Calorimetry (DSC) thermogram with an endothermhaving an onset at about 207° C. and a peak at about 220° C. In someembodiments, Compound A-HCl is crystalline and has a DifferentialScanning Calorimetry (DSC) thermogram substantially the same as shown inFIG. 2(a). In some embodiments, Compound A-HCl is crystalline and has aninfrared (IR) spectrum with characteristic peaks at 2223 cm⁻¹, 1620cm⁻¹, 1595 cm⁻¹, 1457 cm⁻¹, 1238 cm⁻¹, 1220 cm⁻¹, and 1117 cm⁻¹. In someembodiments, Compound A-HCl is crystalline and has an infrared (IR)spectrum substantially the same as shown in FIG. 3. In some embodiments,Compound A-HCl is crystalline and has reversible water uptake (˜4.5%w/w) between 2 and 95% Relative Humidity (RH). In some embodiments,Compound A-HCl is crystalline and has reversible water uptake (˜2.3%)between 15 and 75% Relative Humidity (RH). In some embodiments, CompoundA-HCl is crystalline and has an unchanged XRPD after DVS analysis at 90%RH and room temperature over 24 hours. In some embodiments, CompoundA-HCl is crystalline and has an unchanged XRPD after the DVS analysis at75% RH and 40° C. over one week.

In some embodiments, provided herein is a pharmaceutically acceptablesalt of Compound A. In some embodiments, the pharmaceutically acceptablesalt of Compound A is the dihydrochloride salt (Compound A-2HCl):

In some embodiments, Compound A-2HCl is amorphous.

In some embodiments, Compound A-2HCl is crystalline.

In some embodiments, Compound A-2HCl is crystalline and is characterizedas having: an X-ray powder diffraction (XRPD) pattern with peaks at 5.4°2-Theta, and 7.3° 2-Theta; an X-ray powder diffraction (XRPD) patternsubstantially the same as shown in FIG. 4; a Differential ScanningCalorimetry (DSC) thermogram with an endotherm having an onset at about233° C. and a peak at about 252° C.; a Differential Scanning Calorimetry(DSC) thermogram substantially the same as shown in FIG. 5(a); aThermogravimetric Analysis (TGA) thermogram substantially the same asshown in FIG. 5(b); an infrared (IR) spectrum with characteristic peaksat 2227 cm⁻¹, 1620 cm⁻¹, 1594 cm⁻¹, 1456 cm⁻¹, 1439 cm⁻¹, 1321 cm⁻¹, and1122 cm⁻¹; an infrared (IR) spectrum substantially the same as shown inFIG. 6; reversible water uptake (˜18% w/w) between 2 and 95% RelativeHumidity (RH); reversible water uptake (˜9% w/w) between 2 and 95%Relative Humidity (RH); an unchanged XRPD after DVS analysis at 90% RHand room temperature over 24 hours; an unchanged XRPD after the DVSanalysis at 75% RH and 40° C. over one week; or combinations thereof.

In some embodiments, Compound A-2HCl is crystalline and has at least oneof the following properties:

-   -   a) an X-ray powder diffraction (XRPD) pattern with peaks at 5.4°        2-Theta, and 7.3° 2-Theta;    -   b) an X-ray powder diffraction (XRPD) pattern substantially the        same as shown in FIG. 4;    -   c) a Differential Scanning Calorimetry (DSC) thermogram with an        endotherm having an onset at about 233° C. and a peak at about        252° C.;    -   d) a Differential Scanning Calorimetry (DSC) thermogram        substantially the same as shown in FIG. 5(a);    -   e) an infrared (IR) spectrum with characteristic peaks at 2227        cm⁻¹, 1620 cm⁻¹, 1594 cm⁻¹, 1456 cm⁻¹, 1439 cm⁻¹, 1321 cm⁻¹, and        1122 cm⁻¹;    -   f) an infrared (IR) spectrum substantially the same as shown in        FIG. 6;    -   g) reversible water uptake (˜18% w/w) between 2 and 95% Relative        Humidity (RH);    -   h) reversible water uptake (˜9% w/w) between 2 and 95% Relative        Humidity (RH);    -   i) an unchanged XRPD after DVS analysis at 90% RH and room        temperature over 24 hours; or    -   j) an unchanged XRPD after the DVS analysis at 75% RH and 40° C.        over one week.

In some embodiments, Compound A-2HCl is crystalline and is characterizedas having at least two of the properties selected from a) to j). In someembodiments, Compound A-2HCl is crystalline and is characterized ashaving at least three of the properties selected from a) to j). In someembodiments, Compound A-2HCl is crystalline and is characterized ashaving at least four of the properties selected from a) to j). In someembodiments, Compound A-2HCl is crystalline and is characterized ashaving at least five of the properties selected from a) to j). In someembodiments, Compound A-2HCl is crystalline and is characterized ashaving at least six of the properties selected from a) to j). In someembodiments, Compound A-2HCl is crystalline and is characterized ashaving at least seven of the properties selected from a) to j). In someembodiments, Compound A-2HCl is crystalline and is characterized ashaving at least eight of the properties selected from a) to j). In someembodiments, Compound A-2HCl is crystalline and is characterized ashaving at least nine of the properties selected from a) to j). In someembodiments, Compound A-2HCl is crystalline and is characterized ashaving properties a), b), c), d), e), f), g), h), i), and j).

In some embodiments, Compound A-2HCl is crystalline and has an X-raypowder diffraction (XRPD) pattern with characteristic peaks at 5.4°2-Theta, and 7.3° 2-Theta. In some embodiments, Compound A-2HCl iscrystalline and has an X-ray powder diffraction (XRPD) patternsubstantially the same as shown in FIG. 4. In some embodiments, CompoundA-2HCl is crystalline and has a Differential Scanning Calorimetry (DSC)thermogram with an endotherm having an onset at about 233° C. and a peakat about 252° C. In some embodiments, Compound A-2HCl is crystalline andhas a Differential Scanning Calorimetry (DSC) thermogram substantiallythe same as shown in FIG. 5(a). In some embodiments, Compound A-2HCl iscrystalline and has an infrared (IR) spectrum with characteristic peaksat 2227 cm⁻¹, 1620 cm⁻¹, 1594 cm⁻¹, 1456 cm⁻¹, 1439 cm⁻¹, 1321 cm⁻¹, and1122 cm⁻¹. In some embodiments, Compound A-2HCl is crystalline and hasan infrared (IR) spectrum substantially the same as shown in FIG. 6. Insome embodiments, Compound A-2HCl is crystalline and has reversiblewater uptake (˜18% w/w) between 2 and 95% Relative Humidity (RH). Insome embodiments, Compound A-2HCl is crystalline and has reversiblewater uptake (˜9% w/w) between 2 and 95% Relative Humidity (RH). In someembodiments, Compound A-2HCl is crystalline and has an unchanged XRPDafter DVS analysis at 90% RH and room temperature over 24 hours. In someembodiments, Compound A-2HCl is crystalline and has an unchanged XRPDafter the DVS analysis at 75% RH and 40° C. over one week.

Synthesis of Compound A, Compound A-HCl, and Compound A-2HCl

Compounds described herein are synthesized using standard synthetictechniques or using methods known in the art in combination with methodsdescribed herein. Unless otherwise indicated, conventional methods ofmass spectroscopy, NMR, HPLC are employed.

Compounds are prepared using standard organic chemistry techniques suchas those described in, for example, March's Advanced Organic Chemistry,6^(th) Edition, John Wiley and Sons, Inc. Alternative reactionconditions for the synthetic transformations described herein may beemployed such as variation of solvent, reaction temperature, reactiontime, as well as different chemical reagents and other reactionconditions.

In the reactions described, it may be necessary to protect reactivefunctional groups, for example hydroxy or amino groups, where these aredesired in the final product, in order to avoid their unwantedparticipation in reactions. A detailed description of techniquesapplicable to the creation of protecting groups and their removal aredescribed in Greene and Wuts, Protective Groups in Organic Synthesis,3rd Ed., John Wiley & Sons, New York, N.Y., 1999, and Kocienski,Protective Groups, Thieme Verlag, New York, N.Y., 1994, which areincorporated herein by reference for such disclosure).

Disclosed herein are methods for the synthesis of Compound A, CompoundA-HCl, and Compound A-2HCl, as outlined in Scheme A.

In some embodiments, chlorination of Compound A-I yields compound A-II.Compound A-II is reacted with a brominating or chlorinating agent toprovide Compound A-IIIa (wherein X is Cl or Br). In some embodiments,Compound A-II is reacted with a brominating agent to provide CompoundA-IIIa, where X is Br (i.e. Compound A-III). Coupling A-IIIa with4-(N-PG-amino)piperidine yields compound A-IVa (where PG is a suitableamino protecting group). Compound A-IVa undergoes a Suzuki reaction withcompound 1 to yield Compound A-Va, which undergoes a second Suzukireaction with (3,5-difluorophenyl)B (where each B is independently aboronic acid, boronate ester, or trifluoroborate) to give CompoundA-VIa. In some embodiments, each B is a boronic acid. In someembodiments, (3,5-difluorophenyl)B is (3,5-difluorophenyl)boronic acid.In some embodiments, the same Pd catalyst is used in both Suzukireactions. In other embodiments, different Pd catalysts are used in eachSuzuki reaction. In some embodiments, each Suzuki reaction is performedin the same solvent. In other embodiments, each Suzuki reaction isperformed in different solvents. In some embodiments, Compound A-Va isisolated prior to the second Suzuki reactions. In other embodiments, thetwo Suzuki reactions are performed in one reaction vessel withoutisolation or purification of Compound A-Va. In some embodiments,residual Palladium is removed from Compound A-VIa via a palladiumscavenger, such as SiO₂, charcoal, L-cysteine, SilicaBond Cysteine,Si-Thiol, SilicaBond DMT, or the like.

Deprotection of the PG group of Compound A-VIa and treatment with HClyields Compound A-2HCl, which is converted to Compound A-HCl. In someembodiments, Compound A-HCl or Compound A-2HCl is treated with anappropriate base, such as sodium hydroxide, sodium carbonate, sodiumbicarbonate, or the like, in order to yield Compound A (the free baseform).

In some embodiments, when PG is tert-butyloxycarbonyl (Boc) thencompound A-VIa is treated with HCl to provide Compound A-2HCl, which isconverted to Compound A-HCl. In some embodiments, Compound A-HCl orCompound A-2HCl is treated with an appropriate base, such as sodiumhydroxide, sodium carbonate, sodium bicarbonate, or the like, in orderto yield Compound A (the free base form).

In some embodiments, the crystalline form of Compound A, Compound A-HCl,or Compound A-2HCl is isolated directly from the reaction mixture. Insome embodiments, the crystalline form of Compound A, Compound A-HCl, orCompound A-2HCl is formed by recrystallization from a suitable solvent.Suitable solvents include, but are not limited to, water, methanol,ethanol, ethyl acetate, isopropyl acetate, pentane, hexanes, heptane, orcombinations thereof.

Synthesis of 6-bromo-3-chloroquinolin-4-ol (A-II)

In some embodiments, Compound A-II is produced by chlorination of A-I.In some embodiments, chlorination is accomplished by treating A-I with achlorinating agent, such as N-chlorosuccinimide (NCS),trichloroisocyanuric acid (TCA), sulfuryl chloride, chlorine, sodiumhypochlorite, calcium hypochlorite, hypochlorous acid, or2,3,4,5,6,6-hexachloro-2,4-cyclohexadien-1-one, or the like. In someembodiments, chlorination is accomplished by treating A-I withN-chlorosuccinimide (NCS).

In some embodiments, chlorination is performed in a suitable solventsuch as acetic acid, water, ethanol, methanol, toluene, dichloromethane,tetrahydrofuran, dioxane, N,N-dimethylformamide, or the like.

In some embodiments, chlorination is performed in acetic acid. In someembodiments, 1 volume of acetic acid refers to an amount that is thesame volume as the reagents in the reaction, i.e., A-I and/or NCS. Insome embodiments, the reaction is performed in about 5, 7.5, 10, 12.5,15, or 20 volumes of acetic acid. In some embodiments, the reaction isperformed in about 10 volumes of acetic acid. In some embodiments, thereaction is performed in about 12.5 volumes of acetic acid. In someembodiments, the reaction is performed in about 20 volumes of aceticacid.

In some embodiments, chlorination is performed at elevated temperature.In some embodiments, the reaction temperature is between about 40° C.and about 150° C. In some embodiments, the reaction temperature isbetween about 40° C. and about 120° C. In some embodiments, the reactiontemperature is between about 40° C. and about 100° C. In someembodiments, the reaction temperature is between about 40° C. and about80° C. In some embodiments, the reaction temperature is between about40° C. and about 60° C. In some embodiments, the reaction temperature isbetween about 45° C. and about 55° C. In some embodiments, the reactiontemperature is about 40° C., 45° C., 50° C., 55° C., or 60° C. In someembodiments the reaction temperature is about 50° C.

Synthesis of 4,6-dibromo-3-chloroquinoline (A-III)

In some embodiments, the hydroxyl group of Compound A-II is converted toa halogen atom (X is Br or Cl). In some embodiments, halogenation isbromination. In some embodiments, halogenation is chlorination.

In some embodiments, Compound A-II is brominated with phosphorustribromide (PBr₃), phosphorus oxybromide (POBr₃), hydrobromic acid,bromine, dibromotriphenylphosphorane, or the like. In some embodiment,A-II is brominated with PBr₃. In some embodiments, when A-II isbrominated, the product is A-III.

In some embodiments, Compound A-II is chlorinated with POCl₃, thionylchloride, or the like. In some embodiments, A-II is chlorinated withPOCl₃.

In some embodiments, the halogenation reaction is conducted in asuitable solvent. In some embodiments, the suitable solvent isacetonitrile, water, ethanol, isopropanol, dichloromethane, toluene,N,N-dimethylformamide, acetic acid, acetone, or the like. In someembodiments, the halogenation solvent is DMF. In some embodiments, thereaction temperature is between 0° C. and 30° C. In some embodiments,the reaction temperature is between 0° C. and room temperature. In someembodiments, the reaction starts at 0° C. and warms to room temperature.

Synthesis of N-protected 1-(6-bromo-3-chloroquinolin-4-yl)piperidine(A-IVa)

In some embodiments, Compound A-IVa is prepared from Compound A-III. Insome embodiments, Compound A-IVa is made from Compound A-III by reactingCompound A-III with an N-protected 4-amino piperidine in the presence ofa base in a suitable solvent.

In some embodiments, the amino protecting group is a carbamate (such as9-fluorenylmethyl carbamate (Fmoc), t-butyl carbamate (Boc), benzylcarbamate (Cbz), or the like), an amide (such as acetyl,trifluoroacetyl, or the like), phalimide, benzyl, trityl,benzylidineamine, tosyl, or the like. In some embodiments, the aminoprotecting group is a carbamate. In some embodiments, the aminoprotecting group is Boc. In some embodiments, when the protecting groupis Boc, the product is Compound A-IV.

In some embodiments, the base is a non-nucleophilic base. In someembodiments, the base is an amine base. In some embodiments, the base isan amine base such as triethylamine (TEA), diisopropylethylamine(DIPEA), 1,8-diazabicycloundec-7-ene (DBU),1,2,2,6,6-pentamethylpiperidine, tributylamine, or the like. In someembodiments, the base is DIPEA.

In some embodiments, the base is an inorganic base. In some embodiments,the base is a carbonate (MCO₃) or bicarbonate (MHCO₃) base, where M issodium, potassium, or cesium. In some embodiments, the base is aninorganic base such as sodium bicarbonate, Na₂CO₃, K₂CO₃, Cs₂CO₃, or thelike.

In some embodiments, the suitable solvent for the reaction isN,N-dimethylformamide, N,N-dimethylacetamide, dimethylsulfoxide,dichlormethane, chloroform, dioxane, tetrahydrofuran, toluene,acetonitrile, ethanol, isopropanol, or the like. In some embodiments,the solvent for the reaction is dimethylsulfoxide.

In some embodiments, the reaction temperature is between about 0° C. andabout 250° C., between about 50° C. and about 200° C., or between about100° C. and about 180° C. In some embodiments, the reaction temperatureis between about 120° C. and about 150° C. In some embodiments, thereaction temperature is about 100, 105, 110, 115, 120, 125, 130, 135,140, 145, 150, 155, 160, 165, 170, 175, or 180° C.

Alternate Synthesis of N-Protected1-(6-bromo-3-chloroquinolin-4-yl)piperidine (Compound A-IVa)

In some embodiments, Compound A-IVa is prepared from Compound 3. In someembodiments, Compound A-IVa is made from Compound 3 by reacting Compound3 with an N-protected 4-amino piperidine in the presence of a base in asuitable solvent to provide compound 4a, followed by a chlorination ofCompound 4a. In some embodiments, when the protecting group is Boc,Compound 4a is Compound 4.

In some embodiments, the amino protecting group is a carbamate (such as9-fluorenylmethyl carbamate (Fmoc), t-butyl carbamate (Boc), benzylcarbamate (Cbz), or the like), an amide (such as acetyl,trifluoroacetyl, or the like), phalimide, benzyl, trityl,benzylidineamine, tosyl, or the like. In some embodiments, the aminoprotecting group is a carbamate. In some embodiments, the aminoprotecting group is Boc. In some embodiments, when the protecting groupis Boc, the product is Compound A-IV.

In some embodiments, the base is a non-nucleophilic base. In someembodiments, the base is an amine base. In some embodiments, the base isan amine base such as triethylamine (TEA), diisopropylethylamine(DIPEA), 1,8-diazabicycloundec-7-ene (DBU),1,2,2,6,6-pentamethylpiperidine, tributylamine, or the like. In someembodiments, the base is DIPEA. In some embodiments, the base is aninorganic base. In some embodiments, the base is a carbonate (MCO₃) orbicarbonate (MHCO₃) base, where M is sodium, potassium, or cesium. Insome embodiments, the base is an inorganic base such as sodiumbicarbonate, Na₂CO₃, K₂CO₃, Cs₂CO₃ or the like. In some embodiments, thesuitable solvent for the reaction is N,N-dimethylformamide,N,N-dimethylacetamide, dimethylsulfoxide, dichlormethane, chloroform,dioxane, tetrahydrofuran, toluene, acetonitrile, ethanol, isopropanol,or the like. In some embodiments, the solvent for the reaction isN,N-dimethylformamide. In some embodiments, the reaction temperature isbetween about 0° C. and about 250° C., between about 50° C. and about200° C., or between about 100° C. and about 180° C. In some embodiments,the reaction temperature is between about 120° C. and about 150° C. Insome embodiments, the reaction temperature is about 100, 105, 110, 115,120, 125, 130, 135, 140, 145, 150, 155, 160, 165, 170, 175, or 180° C.

In some embodiments, Compound A-IVa is produced by chlorination ofCompound 4a. In some embodiments, chlorination is accomplished bytreating Compound 4a with a chlorinating agent, such asN-chlorosuccinimide (NCS), trichloroisocyanuric acid (TCA), sulfurylchloride, chlorine, sodium hypochlorite, calcium hypochlorite,hypochlorous acid, or 2,3,4,5,6,6-hexachloro-2,4-cyclohexadien-1-one, orthe like. In some embodiments, chlorination is accomplished by treatingCompound 4a with N-chlorosuccinimide (NCS). In some embodiments,chlorination is performed in a suitable solvent such as acetic acid,water, ethanol, methanol, toluene, dichloromethane, tetrahydrofuran,dioxane, N,N-dimethylformamide, or the like. In some embodiments, thechlorination is performed in toluene. In some embodiments, thechlorination is performed at elevated temperature. In some embodiments,the reaction temperature is between about 40° C. and about 150° C. Insome embodiments, the reaction temperature is between about 40° C. andabout 120° C. In some embodiments, the reaction temperature is betweenabout 40° C. and about 100° C. In some embodiments, the reactiontemperature is between about 50° C. and about 80° C. In some embodimentsthe reaction temperature is about 70° C.

Synthesis of (3-cyano-2-hydroxyphenyl)boronic Acid (Compound 1)

Also disclosed herein, is a two step process for the synthesis ofCompound 1, wherein B is a boronic acid or boronate ester, the processcomprises (1) protecting the hydroxyl group of Compound 2 with asuitable protecting group (PG′) to generate Compound 2a; (2) reactingCompound 2a with a borylation agent under suitable reaction conditions;and (3) removal of the protecting group (PG′) to provide Compound 1.

In some embodiments, the suitable protecting group (PG′) ismethoxymethyl (MOM), ethoxyethyl (EE), methoxypropyl (MOP),benzyloxymethyl (BOM), 2-methoxyethoxymethyl (MEM), benzyl (Bn),para-methoxybenzyl (PMB), 2-naphthylmethyl (Nap), methyl (Me), allyl,tetrahydropyranyl (THP), acetyl (Ac), benzoyl (Bz), 2,2,2-trichloroethylcarbonyl (Troc), [2-(trimethylsilyl)ethoxy]methyl, trimethylsilyl (TMS),triethylsilyl (TES), triisopropyl silyl (TIPS), tert-butyldimethylsilyl(TBDMS), or tert-butyldiphenylsilyl (TBDPS).

In some embodiments, the suitable protecting group (PG′) ismethoxymethyl, 2-methoxyethoxymethyl, benzyl, para-methoxybenzyl,methyl, allyl, tetrahydropyranyl, [2-(trimethylsilyl)ethoxy]methyl,trimethylsilyl, triethylsilyl, triisopropyl silyl,tert-butyldimethylsilyl, or tert-butyldiphenylsilyl. In someembodiments, the suitable protecting group (PG′) is methoxymethyl, or2-methoxyethoxymethyl. In some embodiments, the suitable protectinggroup (PG′) is methoxymethyl. In some embodiments, the suitableprotecting group (PG′) is acetyl.

In some embodiments, removal of the protecting group in step (3) isaccomplished with different reagents depending on what the protectinggroup is. In some embodiments, removal of the protecting group in step(3) is accomplished by treatment with hydrochloric acid, hydrobromicacid, acetic acid, trifluoroacetic acid, tosic acid, ZnBr₂, hydrogenover Pd/C, 2,3-dichloro-5,6-dicyano-1,4-benzoquinone (DDQ), borontribromide, boron trichloride, trimethylsilyl iodide, Pd(PPh₃)₄,tetra-n-butylammonium fluoride (TBAF), or HF-pyridine. In someembodiments, removal of the protecting group in step (3) is accomplishedby treatment with hydrochloric acid, hydrobromic acid, acetic acid,trifluoroacetic acid, or tosic acid. In some embodiments, removal of theprotecting group in step (3) is accomplished by treatment withhydrochloric acid.

In some embodiments, the borylation agent is pinacolborane,catecholborane, bis(neopentyl glycolato)diboron, bis(pinacolato)diboron,bis(hexylene glycolato)diboron, bis(catecholato)diboron,tetrahydroxydiboron, trimethoxyboron, triisopropoxyboron,4,4,5,5-tetramethyl-1,3,2-dioxaborolane,4,6,6-trimethyl-1,3,2-dioxaborinane, diisopropylamine borane,bis(neopentyl glycolato)diboron, bis(catecholato)diboron, orbis(pinacolato)diboron. In some embodiments, the suitable reactionconditions of (2) comprise the use of metal halogen exchange reagents.In some embodiments, the suitable reaction conditions of (2) comprisethe use of metal halogen exchange reagents selected from Grignardreagents and alkyl lithium reagents. In some embodiments, the suitablereaction conditions of (2) comprise the use of isopropyl magnesiumchloride in tetrahydrofuran. In some embodiments, the borylation agentis triisopropoxyboron and the suitable reaction conditions of (2)comprise the use of isopropyl magnesium chloride in tetrahydrofuran. Insome embodiments, the suitable reaction conditions of (2) comprise theuse of transition metal mediated reaction conditions. In someembodiments, the suitable reaction conditions of (2) comprise the use ofpalladium metal mediated reaction conditions.

In some embodiments, B is a boronic acid or a boronic ester. In someembodiments, B is

In some embodiments, B is a boronic acid. In some embodiments, B is

In some embodiments, B is a boronic ester. In some embodiments, B is

In some embodiments, the process further comprises an additional step of(4) converting the boronic acid or boronate ester to a trifluoroborate.In such embodiments, B is

In some embodiments, this conversion is achieved by treating the boroncontaining compound with KHF₂.

Synthesis of tert-butyl(1-(6-(3-cyano-2-hydroxyphenyl)-3-(3,5-difluorophenyl)quinolin-4-yl)piperidin-4-yl)carbamate(Compound A-VI)

In some embodiments, Compound A-VIa is synthesized from Compound A-IVa.In some embodiments, Compound A-VIa is prepared by conducting twosuccessive Suzuki reactions of A-IVa. In some embodiments, CompoundA-IVa is reacted in a first Suzuki reaction with Compound 1, a suitablepalladium catalyst, suitable ligand, and a suitable base in a suitablesolvent to yield Compound A-Va. In some embodiments, Compound A-Va isreacted in a second Suzuki reaction with 3,5-difluorophenylboronic acid,a suitable palladium catalyst, suitable ligand, and a suitable base in asuitable solvent to yield Compound A-VIa. In some embodiments, CompoundA-Va is isolated prior to the second Suzuki reaction. In someembodiments, Compound A-Va is not isolated prior to the second Suzukireaction. When PG is Boc, then Compound A-Va is Compound A-V.

In some embodiments, suitable bases in Suzuki reactions include aminebases and inorganic bases. Suitable amine bases for Suzuki reactionsinclude, but are not limited to, triethylamine, diisopropylethylamine,1,2,2,6,6-pentamethylpiperidine, tributylamine,1,8-diazabicycloundec-7-ene (DBU), or the like. Suitable inorganic basesfor Suzuki reactions include, but are not limited to, NaOAc, KOAc,Ba(OH)₂, Li₂CO₃, Na₂CO₃, K₂CO₃, Cs₂CO₃, Na₃PO₄, K₃PO₄, or the like. Insome embodiments, the base in each Suzuki reaction is the same. In someembodiments, the base in each Suzuki reaction is not the same. In someembodiments, about 1, 2, 3, 4, 5, or 6 equivalents of the base is usedin the Suzuki reaction(s). In some embodiments, when Compound A-Va isnot isolated prior to the second Suzuki reaction, no additional base isadded to the reaction.

In some embodiments, the amount of palladium for each Suzuki reaction isless than about 0.05 equiv. In some embodiments, the amount of palladiumfor each Suzuki reaction is from about 0.05 equiv to about 0.2 equiv. Insome embodiments, the amount of palladium for each Suzuki reaction isabout 0.05, 0.1, 0.15, or 0.2 equiv. In some embodiments, the amount ofpalladium for each Suzuki reaction is the same. In some embodiments, theamount of palladium for each Suzuki reaction is not the same.

In some embodiments, a suitable ligand is used for the palladiumcatalyst. In some embodiments, the ligand is a phosphine ligand. In someembodiments, the ligand is an aliphatic phosphine ligand, such astrimethyl phosphine, tricyclohexylphosphine, tri-tert-butyl-phosphine orthe like. In some embodiments, the ligand is an aromatic phosphine, suchas XPhos, SPhos, JohnPhos, Amphos, triphenylphosphine,methyldiphenylphosphine, or the like. In some embodiments, the ligand isa phosphite ligand, such as trimethylphosphite, triphenylphosphite, orthe like. In some embodiments, the ligand is a bis-phosphine ligand,such as diphenylphosphinomethane (dppm), diphenyl phosphinoethane(dppe), 1,1′-bis(diphenylphosphino)ferrocene (dppf), or the like. Insome embodiments, the ligand in each Suzuki reaction is the same. Insome embodiments, the ligand for each Suzuki reaction is not the same.

In some embodiments, the palladium source in one or both Suzukireactions is a Pd(0) source, such as Pd₂(dba)₃, Pd(PPh₃)₄, or the like.In some embodiments, the palladium source in one or both Suzukireactions is a Pd(II) source, such as PdCl₂, Pd(OAc)₂, PdCl₂(PPh₃)₂,PdCl₂(dppf).DCM, PdCl₂(Amphos), or the like. In some embodiments, thepalladium source in each Suzuki reaction is the same. In someembodiments, the palladium source in each Suzuki reaction is different.

In some embodiments, the solvent system used in one or both Suzukireactions is a single solvent. In some embodiments, the solvent systemused in one or both Suzuki reaction is a cosolvent mixture. In someembodiments, the solvent system used in one or both Suzuki reactions istoluene, DMF, acetonitrile, EtOH, IPA, THF, dioxane, water, or mixturesthereof. In some embodiments, the solvent system used in each Suzukireaction is the same. In some embodiments, the solvent system used ineach Suzuki reaction is not the same.

In some embodiments, the temperature used in one or both Suzuki reactionis between about 50° and 150° C., preferably between about 60° C. and120° C. In some embodiments, the temperature used in one or both Suzukireaction is between 80° C. and 85° C. In some embodiments, thetemperature used in one or both Suzuki reaction is between 90° C. and100° C. In some embodiments, each Suzuki reaction is performed at thesame temperature. In some embodiments, each Suzuki reaction is performedat a different temperature.

In some embodiments, Compound A-Va is isolated between reactions. Insome embodiments Compound A-Va is not isolated between reactions. Insome embodiments, each Suzuki reaction is performed in the same reactionvessel. In some embodiments, each Suzuki reaction is performed in thesame reaction vessel without isolation of Compound A-Va. In someembodiments, each Suzuki reaction is performed in the same solvent. Insome embodiments, each Suzuki reaction is performed with the same base.

In one aspect, the two Suzuki reactions are performed successively inthe same reaction vessel without isolation of intermediate A-Va, andwithout removal of the solvent, wherein, after the first reaction iscompleted, the reaction vessel is allowed to cool prior to addition ofthe second boronic acid and then the reaction vessel is heated for thesecond Suzuki reaction. In some embodiments, a second palladium sourceand/or ligand is added with the second boronic acid. In someembodiments, no additional palladium source and/or ligand is added withthe second boronic acid. In some embodiments, additional base is addedto the reaction with the second boronic acid. In some embodiments, noadditional base is added to the reaction with the second boronic acid.

In some embodiments, the progress of one or both Suzuki reactions ismonitored by HPLC or by TLC.

In some embodiments, the solvent system used is 10:1 dioxane:water. Insome embodiments, the base used is K₂CO₃ and 4 equiv of base are used.In some embodiments, the first palladium source is PdCl₂(dppf).CH₂Cl₂.In some embodiments, the second palladium source is PdCl₂(Amphos). Insome embodiments, the first Suzuki reaction proceeds at about 80-85° C.In some embodiments, the second Suzuki reaction proceeds at about90-100° C.

In some embodiments, Compound A-VIa contains a detectable amount ofresidual palladium as an impurity. When PG is Boc, then Compound A-VIais Compound A-VI.

In some embodiments, the isolated product of Compound A-VI contains adetectable amount of unreacted Compound A-V. In some embodiments, asample of Compound A-VI contains a detectable amount of an impurityselected from:

In some embodiments, Compound A-VI is purified by recrystallization. Insome embodiments, Compound A-VI is heated in a suitable solvent orsolvent mixture for an appropriate amount of time. In some embodiments,the purity of Compound A-VI is improved by this process (see Table 1below). In some embodiments, this process of recrystallization/slurryingremoves or reduces the amount of residual palladium in samples ofCompound A-VI.

TABLE 1 HPLC purity of Solid^(a) Crystallization (RC)/ Un-reacted Otherslurry Purity (%) Compound A-V (%) Impurity (%) RC from MTBE 96.22 1.052.72 RC from IPA 97.10 0.68 2.22 RC from EA 95.47 2.18 2.35 RC from MeOH92.96 1.60 1.86 Precipitation from 96.28 1.09 1.86 DCM/pet ether Slurryin ACN 95.61 1.32 2.51 RC from THF/water 93.72 2.27 3.40 Precipitationfrom 94.35 2.17 2.85 DMF/water RT Slurry in Toluene 96.36 0.41 3.24 Hotslurry in Toluene 97.57 0.16 2.28 From DCM/MTBE 96.78 0.80 2.42 Hotslurry from IPAc 98.13 0.00 1.87 RC from THF/pet ether 98.24 0.16 1.59Hot slurry in MeOH 96.30 0.35 3.35 ^(a)The purity of the starting A-VIfor all the RC/slurry studies was 96.18%.

Due to the fact that the synthetic methods described above utilize atransition metal catalyst, purification steps are performed to reducethe amount of palladium in the product. Purification steps to reduce theamount of palladium in a product are conducted so that activepharmaceutical ingredients meet palladium specification guidelines.(“Guideline on the Specification Limits for Residues of Metal Catalysts”European Medicines Agency Pre-authorisation Evaluation of Medicines forHuman Use, London, January 2007, Doc. Ref. CPMP/SWP/QWP/4446/00 corr.).In some embodiments, purification steps to reduce the amount ofpalladium in a product includes, but is not limited to, treatment withsolid trimercaptotriazine (TMT), polystyrene-bound TMT, mercapto-porouspolystyrene-bound TMT, polystyrene-bound ethylenediamine, activatedcarbon, glass bead sponges, Smopex™, silica bound scavengers,thiol-derivatized silica gel, N-acetylcysteine, n-Bu3P, crystallization,extraction, 1-cysteine, n-Bu3P/lactic acid (Garrett et al., Adv. Synth.Catal. 2004, 346, 889-900). In some embodiments, activated carbonincludes but is not limited to DARCO® KB-G, DARCO® KB-WJ. In one aspectsilica bound scavengers include but are not limited to

where

denotes silica gel. In some embodiments, the purification steps toreduce the amount of palladium include the use of activated carbon,derivatized silica gel (e.g., thiol derivatized silica gel), orcombinations thereof.

In some embodiments, A-VI is further treated with a metal scavenger toremove residual palladium. In some embodiments, the metal scavengercomprises SiO2, charcoal, aqueous solution of L-cysteine, a Silicyclemetal scavenger, Si-thiol, SiliaBond DMT or SiliaBond Cysteine. In someembodiments, the scavenger loading (w/w) is 1:3, 1:2, or 1:1. In someembodiments, the metal scavenger is Si-thiol.

In some embodiments, crude A-VI as isolated from the reaction is treatedwith a metal scavenger. In some other embodiments, recrystallized A-VIis treated with a metal scavenger. In some of these embodiments,palladium levels are reduced sufficiently to be undetectable.

In some embodiments, the presence of residual heavy metal (e.g.palladium) impurities is determined by utilizing methods known in theart. In some embodiments, the presence of residual heavy metal (e.g.palladium) impurities is determined by the use of inductively coupledplasma mass spectrometry (ICP-MS). In some embodiments, the presence ofresidual heavy metal (e.g. palladium) impurities is determined by theuse of techniques described in U.S. Pharmacopeia General Chapter <231>Heavy Metals.

Synthesis of3-(4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl)-2-hydroxy-benzonitrile,di-HCl Salt (Compound A-2HCl)

In some embodiments, Compound A-VI is treated with hydrochloric acid ina suitable solvent to yield Compound A-2HCl. In some embodiments, thesuitable solvent is isopropyl alcohol (IPA), methyl tert-butyl ether(MTBE), toluene, ethyl acetate, isopropyl acetate, water, or mixturesthereof. In some embodiments, the suitable solvent is isopropyl alcohol,ethyl acetate, or isopropyl acetate. In some embodiments, the suitablesolvent is IPA.

Preparation of Compound A-HCl from Compound A-2HCl

In some embodiments, Compound A-2HCl is dissolved in a suitable solventand treated with aqueous ammonia to yield Compound A-HCl. In someembodiments, the suitable solvent is IPA, water, methyl acetate orcombinations thereof. In some embodiments, the solvent is water. In someembodiments, the solvent is IPA:water in a ratio of 9:1, 8:2, 7:3, 6:4,or 5:5. In some embodiments, the solvent is methyl acetate:water in aratio of 9:1, 8:2, 7:3, 6:4, or 5:5. In some embodiments, additionalsodium bicarbonate is added to the reaction. In some embodiments thesolvent volume is 5, 10, 15, 20, or more than 20 volumes.

In some embodiments, the aqueous ammonia source is saturated ammoniumchloride (28-30%). In other embodiments, the aqueous ammonia source is25%. In some embodiments, about 1 equivalent of ammonia is added. Insome embodiments, less than 1 equivalent of ammonia is added, e.g., 0.8equiv. In some embodiments, more than one equivalent of ammonia isadded, e.g., 1.25 equiv. In some embodiments, the amount of ammoniaadded is determined by monitoring the pH of the solution. In someembodiments, the pH is adjusted to about 4-6 with the addition ofammonia, bicarbonate, or hydroxide. In some embodiments, the pH isadjusted with the addition of ammonia. In some embodiments, the pH isadjusted to about 4.5 to 4.7 with the addition of ammonia.

In some embodiments, the reaction is heated to about 30, 35, 40, 45, 50,55, or 60° C. In some embodiments, the reaction is heated to about 45°C. In some embodiments, the reaction is heated before addition of theammonia.

In some embodiments, water is added to the reaction mixture whileheating. In some embodiments, water is added to the reaction mixture atthe end of the reaction. In some embodiments, water is added over about2 hours to the reaction mixture.

In some embodiments, the slurry is filtered to isolate Compound A-HCl,or solvate thereof.

In some other embodiments, no ammonia is needed to convert CompoundA-2HCl to Compound A-HCl. In some embodiments, Compound A-2HCl isstirred in about 10, about 20, about 30, about 40, or about 50 volumesof water. In some embodiments, Compound A-2HCl is stirred in about 20 toabout 50 volumes of water. In some embodiments, sodium bicarbonate isadded to the water mixture to adjust the pH. In some embodiments,Compound A-HCl is isolated from filtering the solids from this watermixture.

Preparation of Compound A from Compound A-2HCl

In some embodiments, the free base of Compound A is made by treatingCompound A-2HCl with a suitable base in a suitable solvent. In someembodiments, the suitable base is sodium hydroxide, sodium bicarbonate,or the like. In some embodiments, the suitable solvent is water. In someembodiments, the solid is filtered from the mixture to isolate the freebase of Compound A.

Preparation of Compound A-HCl from Compound A

In some embodiments Compound A-HCl is generated by treating the freebase with about 1 equivalent of HCl in a suitable solvent. In someembodiments, the suitable solvent is methanol, ethanol, isopropylalcohol, acetone, methyl acetate, ethyl acetate, isopropyl acetate,tetrahydrofuran, tetrahydropyran, water, acetone, or combinationsthereof. In some embodiments, the HCl source is HCl in IPA, HCl intoluene, HCl in MTBE, or HCl in water. In some embodiments, CompoundA-HCl is isolated by filtering the solids from the reaction mixture.

Additional Compounds

In another aspect, described herein is a compound of Formula (I), or apharmaceutically acceptable salt, pharmaceutically acceptable solvate,diastereomeric mixture, or enantiomer thereof:

wherein:

-   R^(a) is F, Cl, or —CH₃;-   R^(b) is hydrogen, F, Cl, —CH₃, —CN, —OH, or —OCH₃;-   R^(B) is an unsubstituted or substituted phenyl or an unsubstituted    or substituted pyridinyl, wherein if R^(B) is substituted then R^(B)    is substituted with R^(c) and R^(d);-   R^(c) is hydrogen, F, Cl, Br, —CH₃, —CN, —OH, —OCH₃, —C(═NOCH₃)H, or    —C(═NOH)H;-   R^(d) is —OH, or —NH₂; and-   each R^(f) is hydrogen or F.

In some embodiments, the compound of Formula (I) has the structure ofFormula (Ia), or a pharmaceutically acceptable salt, solvate,diastereomeric mixture, individual enantiomers or prodrug thereof:

In some embodiments, R^(B) as described in Table 2.

In some embodiments, compounds described herein have the followingstructure, or a pharmaceutically acceptable salt, solvate,diastereomeric mixture, or individual enantiomers thereof:

wherein,

R^(a), R^(b), and R^(B) are as described in Table 2.

In some embodiments, R^(a), R^(b), and R^(B) are as described in Table2.

Any combination of the groups described above for the various variablesis contemplated herein. Throughout the specification, groups andsubstituents thereof are chosen by one skilled in the field to providestable moieties and compounds.

Exemplary compounds of Formula (I) include the compounds described inTable 2.

TABLE 2

Cpd No. R^(f) R^(B) R^(a) R^(b) 1-1 H

F F 1-2 H

F —CH3 1-3 H

Cl F 1-4 H

F —CH3 1-5 H

Cl F 1-6 H

F F 1-7 H

F H 1-8 H

F H

Compounds in Table 2 are named:

-   1-1:    2-{4-[(4aS,8aS)-octahydro-1H-pyrido[3,4-b][1,4]oxazin-6-yl]-3-(3,5-difluorophenyl)quinolin-6-yl}-6-[(1E)-(hydroxyimino)methyl]phenol;-   1-2:    2-{4-[(4aS,8aS)-octahydro-1H-pyrido[3,4-b][1,4]oxazin-6-yl]-3-(3-fluoro-5-methylphenyl)quinolin-6-yl}-4-[(1E)-(methoxyimino)methyl]pyridin-3-amine;-   1-3:    2-{4-[(4aS,8aS)-octahydro-1H-pyrido[3,4-b][1,4]oxazin-6-yl]-3-(3-chloro-5-fluorophenyl)quinolin-6-yl}-4-[(1E)-(methoxyimino)methyl]pyridin-3-amine;-   1-4:    2-{4-[(4aS,8aS)-octahydro-1H-pyrido[3,4-b][1,4]oxazin-6-yl]-3-(3-fluoro-5-methylphenyl)quinolin-6-yl}-3-aminopyridine-4-carbonitrile;-   1-5:    2-{4-[(4aS,8aS)-octahydro-1H-pyrido[3,4-b][1,4]oxazin-6-yl]-3-(3-chloro-5-fluorophenyl)quinolin-6-yl}-3-aminopyridine-4-carbonitrile;-   1-6:    2-{4-[(4aS,8aS)-octahydro-1H-pyrido[3,4-b][1,4]oxazin-6-yl]-3-(3,5-difluorophenyl)quinolin-6-yl}-4-[(1E)-(hydroxyimino)methyl]pyridin-3-amine;-   1-7:    2-{4-[(4aS,8aS)-octahydro-1H-pyrido[3,4-b][1,4]oxazin-6-yl]-3-(3-fluorophenyl)quinolin-6-yl}-3-aminopyridine-4-carbonitrile;    and-   1-8:    2-{4-[(4aS,8aS)-octahydro-1H-pyrido[3,4-b][1,4]oxazin-6-yl]-3-(3-fluorophenyl)quinolin-6-yl}-4-methylpyridin-3-amine.

In some embodiments, provided herein is a pharmaceutically acceptablesalt of a compound that is described in Table 2.

Also provided herein is 1-9:3-[4-(4-aminopiperidin-1-yl)-5-fluoro-3-(3-fluoro-5-methylphenyl)quinolin-6-yl]-2-hydroxybenzonitrile,or a pharmaceutically acceptable salt thereof.

Also provided herein is 1-10:3-[4-(4-aminopiperidin-1-yl)-7-fluoro-3-(3-fluoro-5-methylphenyl)quinolin-6-yl]-2-hydroxybenzonitrile,or a pharmaceutically acceptable salt thereof.

In one aspect, compounds described herein are in the form ofpharmaceutically acceptable salts. As well, active metabolites of thesecompounds having the same type of activity are included in the scope ofthe present disclosure. In addition, the compounds described herein canexist in unsolvated as well as solvated forms with pharmaceuticallyacceptable solvents such as water, ethanol, and the like. The solvatedforms of the compounds presented herein are also considered to bedisclosed herein.

In some embodiments, compounds described herein are prepared asdescribed in Scheme B.

In some embodiments, Compound I is converted to Compound II by anucleophilic replacement with a corresponding cyclic amine over chloroin the presence of Et₃N or DIEA. Selective Suzuki-Miyaura reactions withR^(B)B(OH)₂ or its boronic ester yield Compound IV, which can also beprepared by a two-step sequence through the formation of boronic ester(Compound III) followed by selective Suzuki-Miyaura reactions withR^(B)X (X=Cl, Br, or I). Suzuki-Miyaura reactions with R^(A)(OH)₂ or itsboronic ester followed by removal of all protecting groups usingappropriate de-protection methods such as acid lead to the formation ofCompound V.

In some embodiments, the compounds obtained from the above mentionedmethods are prepared as racemic or diastereomic mixtures. In some otherembodiments, racemic mixtures of the compounds are separated to obtainoptically pure (or optically enriched) isomers by the use of commonchiral separation methods such as chiral HPLC, chiral supercriticalfluid chromatographic system (SFC), simulated moving bed chromatography(SMB), and the like.

In some other embodiments, diastereomic mixtures of the compounds areseparated to obtain optically pure (or optically enriched) isomers bythe use of crystallization methods or common non-chiral chromatographymethods such as silica gel chromatography or chiral chromatographymethods such as chiral HPLC, chiral supercritical fluid chromatographicsystem (SFC), simulated moving bed chromatography (SMB), and the like.

In some embodiments, compounds described herein are synthesized asoutlined in the Examples.

“Pharmaceutically acceptable,” as used herein, refers a material, suchas a carrier or diluent, which does not abrogate the biological activityor properties of the compound, and is relatively nontoxic, i.e., thematerial is administered to an individual without causing undesirablebiological effects or interacting in a deleterious manner with any ofthe components of the composition in which it is contained.

The term “pharmaceutically acceptable salt” refers to a form of atherapeutically active agent that consists of a cationic form of thetherapeutically active agent in combination with a suitable anion, or inalternative embodiments, an anionic form of the therapeutically activeagent in combination with a suitable cation. Handbook of PharmaceuticalSalts: Properties, Selection and Use. International Union of Pure andApplied Chemistry, Wiley-VCH 2002. S. M. Berge, L. D. Bighley, D. C.Monkhouse, J. Pharm. Sci. 1977, 66, 1-19. P. H. Stahl and C. G. Wermuth,editors, Handbook of Pharmaceutical Salts: Properties, Selection andUse, Weinheim/Zurich: Wiley-VCH/VHCA, 2002. Pharmaceutical saltstypically are more soluble and more rapidly soluble in stomach andintestinal juices than non-ionic species and so are useful in soliddosage forms. Furthermore, because their solubility often is a functionof pH, selective dissolution in one or another part of the digestivetract is possible and this capability can be manipulated as one aspectof delayed and sustained release behaviours. Also, because thesalt-forming molecule can be in equilibrium with a neutral form, passagethrough biological membranes can be adjusted.

In some embodiments, pharmaceutically acceptable salts are obtained byreacting a compound disclosed herein with an acid. In some embodiments,the compound disclosed herein (i.e. free base form) is basic and isreacted with an organic acid or an inorganic acid. Inorganic acidsinclude, but are not limited to, hydrochloric acid, hydrobromic acid,sulfuric acid, phosphoric acid, nitric acid, and metaphosphoric acid.Organic acids include, but are not limited to, 1-hydroxy-2-naphthoicacid; 2,2-dichloroacetic acid; 2-hydroxyethanesulfonic acid;2-oxoglutaric acid; 4-acetamidobenzoic acid; 4-aminosalicylic acid;acetic acid; adipic acid; ascorbic acid (L); aspartic acid (L);benzenesulfonic acid; benzoic acid; camphoric acid (+);camphor-10-sulfonic acid (+); capric acid (decanoic acid); caproic acid(hexanoic acid); caprylic acid (octanoic acid); carbonic acid; cinnamicacid; citric acid; cyclamic acid; dodecylsulfuric acid;ethane-1,2-disulfonic acid; ethanesulfonic acid; formic acid; fumaricacid; galactaric acid; gentisic acid; glucoheptonic acid (D); gluconicacid (D); glucuronic acid (D); glutamic acid; glutaric acid;glycerophosphoric acid; glycolic acid; hippuric acid; isobutyric acid;lactic acid (DL); lactobionic acid; lauric acid; maleic acid; malic acid(−L); malonic acid; mandelic acid (DL); methanesulfonic acid;naphthalene-1,5-disulfonic acid; naphthalene-2-sulfonic acid; nicotinicacid; oleic acid; oxalic acid; palmitic acid; pamoic acid; phosphoricacid; proprionic acid; pyroglutamic acid (−L); salicylic acid; sebacicacid; stearic acid; succinic acid; sulfuric acid; tartaric acid (+L);thiocyanic acid; toluenesulfonic acid (p); and undecylenic acid.

In some embodiments, a compound disclosed herein is prepared as achloride salt, sulfate salt, bromide salt, mesylate salt, maleate salt,citrate salt or phosphate salt.

In some embodiments, pharmaceutically acceptable salts are obtained byreacting a compound disclosed herein with a base. In some embodiments,the compound disclosed herein is acidic and is reacted with a base. Insuch situations, an acidic proton of the compound disclosed herein isreplaced by a metal ion, e.g., lithium, sodium, potassium, magnesium,calcium, or an aluminum ion. In some cases, compounds described hereincoordinate with an organic base, such as, but not limited to,ethanolamine, diethanolamine, triethanolamine, tromethamine, meglumine,N-methylglucamine, dicyclohexylamine, tris(hydroxymethyl)methylamine. Inother cases, compounds described herein form salts with amino acids suchas, but not limited to, arginine, lysine, and the like. Acceptableinorganic bases used to form salts with compounds that include an acidicproton, include, but are not limited to, aluminum hydroxide, calciumhydroxide, potassium hydroxide, sodium carbonate, potassium carbonate,sodium hydroxide, lithium hydroxide, and the like. In some embodiments,the compounds provided herein are prepared as a sodium salt, calciumsalt, potassium salt, magnesium salt, meglumine salt, N-methylglucaminesalt or ammonium salt.

It should be understood that a reference to a pharmaceuticallyacceptable salt includes the solvent addition forms. In someembodiments, solvates contain either stoichiometric ornon-stoichiometric amounts of a solvent, and are formed during theprocess of crystallization with pharmaceutically acceptable solventssuch as water, ethanol, and the like. Hydrates are formed when thesolvent is water, or alcoholates are formed when the solvent is alcohol.Solvates of compounds described herein are conveniently prepared orformed during the processes described herein. In addition, the compoundsprovided herein optionally exist in unsolvated as well as solvatedforms.

Therapeutic agents that are administrable to mammals, such as humans,must be prepared by following regulatory guidelines. Such governmentregulated guidelines are referred to as Good Manufacturing Practice(GMP). GMP guidelines outline acceptable contamination levels of activetherapeutic agents, such as, for example, the amount of residual solventin the final product. Preferred solvents are those that are suitable foruse in GMP facilities and consistent with industrial safety concerns.Categories of solvents are defined in, for example, the InternationalConference on Harmonization of Technical Requirements for Registrationof Pharmaceuticals for Human Use (ICH), “Impurities: Guidelines forResidual Solvents, Q3C(R3), (November 2005).

Solvents are categorized into three classes. Class 1 solvents are toxicand are to be avoided. Class 2 solvents are solvents to be limited inuse during the manufacture of the therapeutic agent. Class 3 solventsare solvents with low toxic potential and of lower risk to human health.Data for Class 3 solvents indicate that they are less toxic in acute orshort-term studies and negative in genotoxicity studies.

Class 1 solvents, which are to be avoided, include: benzene; carbontetrachloride; 1,2-dichloroethane; 1,1-dichloroethene; and1,1,1-trichloroethane.

Examples of Class 2 solvents are: acetonitrile, chlorobenzene,chloroform, cyclohexane, 1,2-dichloroethene, dichloromethane,1,2-dimethoxyethane, N,N-dimethylacetamide, N,N-dimethylformamide,1,4-dioxane, 2-ethoxyethanol, ethyleneglycol, formamide, hexane,methanol, 2-methoxyethanol, methylbutyl ketone, methylcyclohexane,N-methylpyrrolidine, nitromethane, pyridine, sulfolane, tetralin,toluene, 1,1,2-trichloroethene and xylene.

Class 3 solvents, which possess low toxicity, include: acetic acid,acetone, anisole, 1-butanol, 2-butanol, butyl acetate, tert-butylmethylether (MTBE), cumene, dimethyl sulfoxide, ethanol, ethyl acetate, ethylether, ethyl formate, formic acid, heptane, isobutyl acetate, isopropylacetate, methyl acetate, 3-methyl-1-butanol, methylethyl ketone,methylisobutyl ketone, 2-methyl-1-propanol, pentane, 1-pentanol,1-propanol, 2-propanol, propyl acetate, and tetrahydrofuran.

Residual solvents in active pharmaceutical ingredients (APIs) originatefrom the manufacture of API. In some cases, the solvents are notcompletely removed by practical manufacturing techniques. Appropriateselection of the solvent for the synthesis of APIs may enhance theyield, or determine characteristics such as crystal form, purity, andsolubility. Therefore, the solvent is a critical parameter in thesynthetic process.

In some embodiments, compositions comprising Compound A, or apharmaceutically acceptable salt thereof, comprise an organicsolvent(s). In some embodiments, compositions comprising Compound A, ora pharmaceutically acceptable salt thereof, include a residual amount ofan organic solvent(s). In some embodiments, compositions comprisingCompound A, or a pharmaceutically acceptable salt thereof, comprise aresidual amount of a Class 3 solvent. In some embodiments, the Class 3solvent is selected from the group consisting of acetic acid, acetone,anisole, 1-butanol, 2-butanol, butyl acetate, tert-butylmethyl ether,cumene, dimethyl sulfoxide, ethanol, ethyl acetate, ethyl ether, ethylformate, formic acid, heptane, isobutyl acetate, isopropyl acetate,methyl acetate, 3-methyl-1-butanol, methylethyl ketone, methylisobutylketone, 2-methyl-1-propanol, pentane, 1-pentanol, 1-propanol,2-propanol, propyl acetate, and tetrahydrofuran. In some embodiments,the Class 3 solvent is selected from ethyl acetate, isopropyl acetate,tert-butylmethylether, heptane, isopropanol, and ethanol.

In some embodiments, the compositions comprising Compound A, or apharmaceutically acceptable salt thereof, include a detectable amount ofan organic solvent. In some embodiments, the pharmaceutically acceptablesalt of Compound A is an HCl salt (i.e., Compound A-HCl). In someembodiments, the organic solvent is a Class 3 solvent.

In other embodiments are compositions comprising Compound A, or apharmaceutically acceptable salt thereof, wherein the compositioncomprises a detectable amount of solvent that is less than about 1%,wherein the solvent is selected from acetone, 1,2-dimethoxyethane,acetonitrile, ethyl acetate, tetrahydrofuran, methanol, ethanol,heptane, and 2-propanol. In a further embodiment are compositionscomprising Compound A, or a pharmaceutically acceptable salt thereof,wherein the composition comprises a detectable amount of solvent whichis less than about 5000 ppm. In yet a further embodiment arecompositions comprising Compound A, wherein the detectable amount ofsolvent is less than about 5000 ppm, less than about 4000 ppm, less thanabout 3000 ppm, less than about 2000 ppm, less than about 1000 ppm, lessthan about 500 ppm, or less than about 100 ppm.

The methods and formulations described herein include the use ofN-oxides (if appropriate), or pharmaceutically acceptable salts ofcompounds having the structure disclosed herein, as well as activemetabolites of these compounds having the same type of activity.

In some embodiments, sites on the organic radicals (e.g. alkyl groups,aromatic rings) of compounds disclosed herein are susceptible to variousmetabolic reactions. Incorporation of appropriate substituents on theorganic radicals will reduce, minimize or eliminate this metabolicpathway. In specific embodiments, the appropriate substituent todecrease or eliminate the susceptibility of the aromatic ring tometabolic reactions is, by way of example only, a halogen, deuterium, analkyl group, a haloalkyl group, or a deuteroalkyl group.

In another embodiment, the compounds described herein are labeledisotopically (e.g. with a radioisotope) or by another other means,including, but not limited to, the use of chromophores or fluorescentmoieties, bioluminescent labels, or chemiluminescent labels.

Compounds described herein include isotopically-labeled compounds, whichare identical to those recited in the various formulae and structurespresented herein, but for the fact that one or more atoms are replacedby an atom having an atomic mass or mass number different from theatomic mass or mass number usually found in nature. Examples of isotopesthat can be incorporated into the present compounds include isotopes ofhydrogen, carbon, nitrogen, oxygen, sulfur, fluorine chlorine, iodine,phosphorus, such as, for example, ²H, ³H, ¹³C, ¹⁴C, ¹⁵N, ¹⁸O, ¹⁷O, ³⁵S,¹⁸F, ³⁶Cl, ¹²³I, ¹²⁴I, ¹²⁵I, ¹³¹I, ³²P and ³³P. In one aspect,isotopically-labeled compounds described herein, for example those intowhich radioactive isotopes such as ³H and ¹⁴C are incorporated, areuseful in drug and/or substrate tissue distribution assays. In oneaspect, substitution with isotopes such as deuterium affords certaintherapeutic advantages resulting from greater metabolic stability, suchas, for example, increased in vivo half-life or altered metabolicpathways to reduce undesirable metabolites or reduced dosagerequirements.

In some embodiments, one or more hydrogen atoms on Compound A arereplaced with deuterium. In some embodiments, substitution withdeuterium affords certain therapeutic advantages resulting from greatermetabolic stability, such as, for example, increased in vivo half-lifeor reduced dosage requirements.

In one aspect, described is a compound with the following structure:

wherein,

each R is independently selected from hydrogen or deutrium,

or a pharmaceutically acceptable salt thereof.

In some embodiments, the pharmaceutically acceptable salt of thecompound is an HCl salt. In some embodiments, the pharmaceuticallyacceptable salt of the compound is a DCl salt.

In some embodiments, the compounds disclosed herein possess one or morestereocenters and each stereocenter exists independently in either the Ror S configuration. For example, in some embodiments, the compounddisclosed herein exists in the R configuration when one stereocenter ispresent. In other embodiments, the compound disclosed herein exists inthe S configuration when one stereocenter is present. In someembodiments, the compound disclosed herein exists in the RRconfiguration when two stereocenters are present. In some embodiments,the compound disclosed herein exists in the RS configuration when twostereocenters are present. In some embodiments, the compound disclosedherein exists in the SS configuration when two stereocenters arepresent. In some embodiments, the compound disclosed herein exists inthe SR configuration when two stereocenters are present.

The compounds presented herein include all diastereomeric, individualenantiomers, atropisomers, and epimeric forms as well as the appropriatemixtures thereof. The compounds and methods provided herein include allcis, trans, syn, anti, entgegen (E), and zusammen (Z) isomers as well asthe appropriate mixtures thereof.

Individual stereoisomers are obtained, if desired, by methods such as,stereoselective synthesis and/or the separation of stereoisomers bychiral chromatographic columns or the separation of diastereomers byeither non-chiral or chiral chromatographic columns or crystallizationand recrystallization in a proper solvent or a mixture of solvents. Incertain embodiments, compounds disclosed herein are prepared as theirindividual stereoisomers by reacting a racemic mixture of the compoundwith an optically active resolving agent to form a pair ofdiastereoisomeric compounds/salts, separating the diastereomers andrecovering the optically pure individual enantiomers. In someembodiments, resolution of individual enantiomers of compounds disclosedherein is carried out using covalent diastereomeric derivatives of thecompounds described herein. In another embodiment, diastereomers ofcompounds disclosed herein are separated by separation/resolutiontechniques based upon differences in solubility. In other embodiments,separation of steroisomers of compounds disclosed herein is performed bychromatography or by the forming diastereomeric salts and separation byrecrystallization, or chromatography, or any combination thereof. JeanJacques, Andre Collet, Samuel H. Wilen, “Enantiomers, Racemates andResolutions”, John Wiley And Sons, Inc., 1981. In some embodiments,stereoisomers are obtained by stereoselective synthesis.

Separation of individual enantiomers from a racemic mixture is possibleby the use of chiral supercritical fluid chromatography (SFC) or chiralhigh performance liquid chromatography (HPLC). In some embodiments,enantiomers described herein are separated from each other by the use ofchiral SFC or chrial HPLC. In some embodiments, compounds disclosedherein that include one or more chiral centers (e.g. compounds disclosedherein that include the moietytrans-octahydro-1H-pyrido[3,4-b]morpholin-6-yl) are separated intoindividual enantiomers using chiral SFC or chrial HPLC. A wide varietyof conditions and suitable columns are available.

Daicel polysaccharide chiral stationary phases (CSPs) are among thecolumns used for chiral SFC separations. In some embodiments, Daicelanalytical immobilised and coated CHIRALPAK and CHIRALCEL HPLC columnscan be used for SFC analysis.

In some embodiments, screening for the suitability of using a SFC columnis performed on the four main immobilised phases (CHIRALPAK IA, IB, ICand ID) and the four main coated columns (CHIRALPAK AD and AS andCHIRALCEL OD and OJ), with varying concentrations of organic modifier. Avariety of column phases are available, including but not limited to ODand OJ, OX and OZ chlorinated phases, and a range of complementarycellulose based CHIRALCEL phases including OA, OB, OC, OF, OG and OK.

Non-limiting examples of chiral selectors contemplated for use in theseparation of enantiomers include amylose tris (3,5-dimethylphenylcarbamate), cellulose tris (3,5-dimethylphenylcarbamate), cellulose tris (3,5-dichlorophenylcarbamate), amylose tris (3-chlorophenylcarbamate),amylose tris (3, 5-dichlorophenylcarbamate), amylose tris (3-chloro,4-methylphenylcarbamate), amylose tris((S)-alpha-methylbenzylcarbamate), amylose tris(5-chloro-2-methylphenylcarbamate), cellulose tris (4-methylbenzoate),cellulose tris (4-chloro-3-methylphenylcarbamate), and cellulose tris(3-chloro-4-methylphenylcarbamate).

Non-limiting examples of chiral columns contemplated for use in theseparation of enantiomers include CHIRALPAK IA SFC, CHIRALPAK AD-H SFC,CHIRALPAK IB SFC, CHIRALCEL OD-H SFC, CHIRALPAK IC SFC, CHIRALPAK IDSFC, CHIRALPAK IE SFC, CHIRALPAK IF SFC, CHIRALPAK AZ-H SFC, CHIRALPAKAS-H SFC, CHIRALPAK AY-H SFC, CHIRALCEL OJ-H SFC, CHIRALCEL OX-H SFC,and CHIRALCEL OZ-H SFC.

In some embodiments, the identity of and placement of substituents onthe compounds described herein help to minimize undesired activity. Forexample, in some embodiments undesired activity includes undesired hERGinhibition. In some embodiments, the presence of a hydroxyl group and anadjacent cyano group on an aromatic ring reduces undesired hERGinhibition significantly as compared to the lack of both groups, thepresence of a hydroxyl group without an adjacent cyano group, or thepresence of a cyano group without an adjacent hydroxyl group. Forexample, in some embodiments significant reduction of undesired hERGinhibition is observed when R^(B) is a substituted or unsubstituted2-hydroxy-3-cyanophenyl.

In additional or further embodiments, the compounds described herein aremetabolized upon administration to an organism in need to produce ametabolite that is then used to produce a desired effect, including adesired therapeutic effect.

A “metabolite” of a compound disclosed herein is a derivative of thatcompound that is formed when the compound is metabolized. The term“active metabolite” refers to a biologically active derivative of acompound that is formed when the compound is metabolized. The term“metabolized,” as used herein, refers to the sum of the processes(including, but not limited to, hydrolysis reactions and reactionscatalyzed by enzymes) by which a particular substance is changed by anorganism. Thus, enzymes may produce specific structural alterations to acompound. For example, cytochrome P450 catalyzes a variety of oxidativeand reductive reactions while uridine diphosphate glucuronyltransferasescatalyze the transfer of an activated glucuronic-acid molecule toaromatic alcohols, aliphatic alcohols, carboxylic acids, amines and freesulphydryl groups. Metabolites of the compounds disclosed herein areoptionally identified either by administration of compounds to a hostand analysis of tissue samples from the host, or by incubation ofcompounds with hepatic cells in vitro and analysis of the resultingcompounds.

Unless otherwise stated, the following terms used in this applicationhave the definitions given below. The use of the term “including” aswell as other forms, such as “include”, “includes,” and “included,” isnot limiting. The section headings used herein are for organizationalpurposes only and are not to be construed as limiting the subject matterdescribed.

The term “halo” or, alternatively, “halogen” or “halide” means fluoro,chloro, bromo or iodo. In some embodiments, halo is fluoro, chloro, orbromo.

The term “bond” or “single bond” refers to a chemical bond between twoatoms, or two moieties when the atoms joined by the bond are consideredto be part of larger substructure. In one aspect, when a group describedherein is a bond, the referenced group is absent thereby allowing a bondto be formed between the remaining identified groups.

The term “moiety” refers to a specific segment or functional group of amolecule. Chemical moieties are often recognized chemical entitiesembedded in or appended to a molecule.

The term “acceptable” with respect to a formulation, composition oringredient, as used herein, means having no persistent detrimentaleffect on the general health of the subject being treated.

The term “modulate” as used herein, means to interact with a targeteither directly or indirectly so as to alter the activity of the target,including, by way of example only, to enhance the activity of thetarget, to inhibit the activity of the target, to limit the activity ofthe target, or to extend the activity of the target.

The term “modulator” as used herein, refers to a molecule that interactswith a target either directly or indirectly. The interactions include,but are not limited to, the interactions of an agonist, partial agonist,an inverse agonist, antagonist, degrader, or combinations thereof. Insome embodiments, a modulator is an agonist.

The terms “administer,” “administering”, “administration,” and the like,as used herein, refer to the methods that may be used to enable deliveryof compounds or compositions to the desired site of biological action.These methods include, but are not limited to oral routes, intraduodenalroutes, parenteral injection (including intravenous, subcutaneous,intraperitoneal, intramuscular, intravascular or infusion), topical andrectal administration. Those of skill in the art are familiar withadministration techniques that can be employed with the compounds andmethods described herein. In some embodiments, the compounds andcompositions described herein are administered orally.

The terms “co-administration” or the like, as used herein, are meant toencompass administration of the selected therapeutic agents to a singlepatient, and are intended to include treatment regimens in which theagents are administered by the same or different route of administrationor at the same or different time.

The terms “effective amount” or “therapeutically effective amount,” asused herein, refer to a sufficient amount of an agent or a compoundbeing administered, which will relieve to some extent one or more of thesymptoms of the disease or condition being treated. The result includesreduction and/or alleviation of the signs, symptoms, or causes of adisease, or any other desired alteration of a biological system. Forexample, an “effective amount” for therapeutic uses is the amount of thecomposition comprising a compound as disclosed herein required toprovide a clinically significant decrease in disease symptoms. Anappropriate “effective” amount in any individual case is optionallydetermined using techniques, such as a dose escalation study.

The terms “enhance” or “enhancing,” as used herein, means to increase orprolong either in potency or duration a desired effect. Thus, in regardto enhancing the effect of therapeutic agents, the term “enhancing”refers to the ability to increase or prolong, either in potency orduration, the effect of other therapeutic agents on a system. An“enhancing-effective amount,” as used herein, refers to an amountadequate to enhance the effect of another therapeutic agent in a desiredsystem.

The term “pharmaceutical combination” as used herein, means a productthat results from the mixing or combining of more than one activeingredient and includes both fixed and non-fixed combinations of theactive ingredients. The term “fixed combination” means that the activeingredients, e.g. a compound disclosed herein, or a pharmaceuticallyacceptable salt thereof, and a co-agent, are both administered to apatient simultaneously in the form of a single entity or dosage. Theterm “non-fixed combination” means that the active ingredients, e.g. acompound disclosed herein, or a pharmaceutically acceptable saltthereof, and a co-agent, are administered to a patient as separateentities either simultaneously, concurrently or sequentially with nospecific intervening time limits, wherein such administration provideseffective levels of the two compounds in the body of the patient. Thelatter also applies to cocktail therapy, e.g. the administration ofthree or more active ingredients.

The terms “article of manufacture” and “kit” are used as synonyms.

The term “subject” or “patient” encompasses mammals. Examples of mammalsinclude, but are not limited to, any member of the Mammalian class:humans, non-human primates such as chimpanzees, and other apes andmonkey species; farm animals such as cattle, horses, sheep, goats,swine; domestic animals such as rabbits, dogs, and cats; laboratoryanimals including rodents, such as rats, mice and guinea pigs, and thelike. In one aspect, the mammal is a human.

The terms “treat,” “treating” or “treatment,” as used herein, includealleviating, abating or ameliorating at least one symptom of a diseaseor condition, preventing additional symptoms, inhibiting the disease orcondition, e.g., arresting the development of the disease or condition,relieving the disease or condition, causing regression of the disease orcondition, relieving a condition caused by the disease or condition, orstopping the symptoms of the disease or condition eitherprophylactically and/or therapeutically.

Pharmaceutical Compositions

In some embodiments, the compounds described herein are formulated intopharmaceutical compositions. Pharmaceutical compositions are formulatedin a conventional manner using one or more pharmaceutically acceptableinactive ingredients that facilitate processing of the active compoundsinto preparations that are used pharmaceutically. Proper formulation isdependent upon the route of administration chosen. A summary ofpharmaceutical compositions described herein is found, for example, inRemington: The Science and Practice of Pharmacy, Nineteenth Ed (Easton,Pa.: Mack Publishing Company, 1995); Hoover, John E., Remington'sPharmaceutical Sciences, Mack Publishing Co., Easton, Pa. 1975;Liberman, H. A. and Lachman, L., Eds., Pharmaceutical Dosage Forms,Marcel Decker, New York, N.Y., 1980; and Pharmaceutical Dosage Forms andDrug Delivery Systems, Seventh Ed. (Lippincott Williams & Wilkins 1999),herein incorporated by reference for such disclosure.

In some embodiments, the compounds described herein are administeredeither alone or in combination with pharmaceutically acceptablecarriers, excipients or diluents, in a pharmaceutical composition.Administration of the compounds and compositions described herein can beeffected by any method that enables delivery of the compounds to thesite of action. These methods include, though are not limited todelivery via enteral routes (including oral, gastric or duodenal feedingtube, rectal suppository and rectal enema), parenteral routes (injectionor infusion, including intraarterial, intracardiac, intradermal,intraduodenal, intramedullary, intramuscular, intraosseous,intraperitoneal, intrathecal, intravascular, intravenous, intravitreal,epidural and subcutaneous), inhalational, transdermal, transmucosal,sublingual, buccal and topical (including epicutaneous, dermal, enema,eye drops, ear drops, intranasal, vaginal) administration, although themost suitable route may depend upon for example the condition anddisorder of the recipient. By way of example only, compounds describedherein can be administered locally to the area in need of treatment, byfor example, local infusion during surgery, topical application such ascreams or ointments, injection, catheter, or implant. The administrationcan also be by direct injection at the site of a diseased tissue ororgan.

In some embodiments, pharmaceutical compositions suitable for oraladministration are presented as discrete units such as capsules, cachetsor tablets each containing a predetermined amount of the activeingredient; as a powder or granules; as a solution or a suspension in anaqueous liquid or a non-aqueous liquid; or as an oil-in-water liquidemulsion or a water-in-oil liquid emulsion. In some embodiments, theactive ingredient is presented as a bolus, electuary or paste.

Pharmaceutical compositions which can be used orally include tablets,push-fit capsules made of gelatin, as well as soft, sealed capsules madeof gelatin and a plasticizer, such as glycerol or sorbitol. Tablets maybe made by compression or molding, optionally with one or more accessoryingredients. Compressed tablets may be prepared by compressing in asuitable machine the active ingredient in a free-flowing form such as apowder or granules, optionally mixed with binders, inert diluents, orlubricating, surface active or dispersing agents. Molded tablets may bemade by molding in a suitable machine a mixture of the powdered compoundmoistened with an inert liquid diluent. In some embodiments, the tabletsare coated or scored and are formulated so as to provide slow orcontrolled release of the active ingredient therein. All formulationsfor oral administration should be in dosages suitable for suchadministration. The push-fit capsules can contain the active ingredientsin admixture with filler such as lactose, binders such as starches,and/or lubricants such as talc or magnesium stearate and, optionally,stabilizers. In soft capsules, the active compounds may be dissolved orsuspended in suitable liquids, such as fatty oils, liquid paraffin, orliquid polyethylene glycols. In some embodiments, stabilizers are added.Dragee cores are provided with suitable coatings. For this purpose,concentrated sugar solutions may be used, which may optionally containgum arabic, talc, polyvinyl pyrrolidone, carbopol gel, polyethyleneglycol, and/or titanium dioxide, lacquer solutions, and suitable organicsolvents or solvent mixtures. Dyestuffs or pigments may be added to thetablets or Dragee coatings for identification or to characterizedifferent combinations of active compound doses.

In some embodiments, pharmaceutical compositions are formulated forparenteral administration by injection, e.g., by bolus injection orcontinuous infusion. Formulations for injection may be presented in unitdosage form, e.g., in ampoules or in multi-dose containers, with anadded preservative. The compositions may take such forms as suspensions,solutions or emulsions in oily or aqueous vehicles, and may containformulatory agents such as suspending, stabilizing and/or dispersingagents. The compositions may be presented in unit-dose or multi-dosecontainers, for example sealed ampoules and vials, and may be stored inpowder form or in a freeze-dried (lyophilized) condition requiring onlythe addition of the sterile liquid carrier, for example, saline orsterile pyrogen-free water, immediately prior to use. Extemporaneousinjection solutions and suspensions may be prepared from sterilepowders, granules and tablets of the kind previously described.

Pharmaceutical compositions for parenteral administration includeaqueous and non-aqueous (oily) sterile injection solutions of the activecompounds which may contain antioxidants, buffers, bacteriostats andsolutes which render the formulation isotonic with the blood of theintended recipient; and aqueous and non-aqueous sterile suspensionswhich may include suspending agents and thickening agents. Suitablelipophilic solvents or vehicles include fatty oils such as sesame oil,or synthetic fatty acid esters, such as ethyl oleate or triglycerides,or liposomes. Aqueous injection suspensions may contain substances whichincrease the viscosity of the suspension, such as sodium carboxymethylcellulose, sorbitol, or dextran. Optionally, the suspension may alsocontain suitable stabilizers or agents which increase the solubility ofthe compounds to allow for the preparation of highly concentratedsolutions.

Pharmaceutical compositions may also be formulated as a depotpreparation. Such long acting formulations may be administered byimplantation (for example subcutaneously or intramuscularly) or byintramuscular injection. Thus, for example, the compounds may beformulated with suitable polymeric or hydrophobic materials (forexample, as an emulsion in an acceptable oil) or ion exchange resins, oras sparingly soluble derivatives, for example, as a sparingly solublesalt.

For buccal or sublingual administration, the compositions may take theform of tablets, lozenges, pastilles, or gels formulated in conventionalmanner. Such compositions may comprise the active ingredient in aflavored basis such as sucrose and acacia or tragacanth.

Pharmaceutical compositions may be administered topically, that is bynon-systemic administration. This includes the application of a compoundof the present invention externally to the epidermis or the buccalcavity and the instillation of such a compound into the ear, eye andnose, such that the compound does not significantly enter the bloodstream. In contrast, systemic administration refers to oral,intravenous, intraperitoneal and intramuscular administration.

Pharmaceutical compositions suitable for topical administration includeliquid or semi-liquid preparations suitable for penetration through theskin to the site of inflammation such as gels, liniments, lotions,creams, ointments or pastes, and drops suitable for administration tothe eye, ear or nose. The active ingredient may comprise, for topicaladministration, from 0.001% to 10% w/w, for instance from 1% to 2% byweight of the formulation.

Pharmaceutical compositions for administration by inhalation areconveniently delivered from an insufflator, nebulizer pressurized packsor other convenient means of delivering an aerosol spray. Pressurizedpacks may comprise a suitable propellant such asdichlorodifluoromethane, trichlorofluoromethane,dichlorotetrafluoroethane, carbon dioxide or other suitable gas. In thecase of a pressurized aerosol, the dosage unit may be determined byproviding a valve to deliver a metered amount. Alternatively, foradministration by inhalation or insufflation, pharmaceuticalpreparations may take the form of a dry powder composition, for examplea powder mix of the compound and a suitable powder base such as lactoseor starch. The powder composition may be presented in unit dosage form,in for example, capsules, cartridges, gelatin or blister packs fromwhich the powder may be administered with the aid of an inhalator orinsufflator.

It should be understood that in addition to the ingredients particularlymentioned above, the compounds and compositions described herein mayinclude other agents conventional in the art having regard to the typeof formulation in question, for example those suitable for oraladministration may include flavoring agents.

Methods of Dosing and Treatment Regimens

In one embodiment, the compounds disclosed herein, or a pharmaceuticallyacceptable salt thereof, are used in the preparation of medicaments forthe treatment of diseases or conditions in a mammal that would benefitfrom modulation of somatostatin activity. Methods for treating any ofthe diseases or conditions described herein in a mammal in need of suchtreatment, involves administration of pharmaceutical compositions thatinclude at least one compound disclosed herein or a pharmaceuticallyacceptable salt, active metabolite, prodrug, or pharmaceuticallyacceptable solvate thereof, in therapeutically effective amounts to saidmammal.

In certain embodiments, the compositions containing the compound(s)described herein are administered for prophylactic and/or therapeutictreatments. In certain therapeutic applications, the compositions areadministered to a patient already suffering from a disease or condition,in an amount sufficient to cure or at least partially arrest at leastone of the symptoms of the disease or condition. Amounts effective forthis use depend on the severity and course of the disease or condition,previous therapy, the patient's health status, weight, and response tothe drugs, and the judgment of the treating physician. Therapeuticallyeffective amounts are optionally determined by methods including, butnot limited to, a dose escalation and/or dose ranging clinical trial.

In prophylactic applications, compositions containing the compoundsdescribed herein are administered to a patient susceptible to orotherwise at risk of a particular disease, disorder or condition. Suchan amount is defined to be a “prophylactically effective amount ordose.” In this use, the precise amounts also depend on the patient'sstate of health, weight, and the like. When used in patients, effectiveamounts for this use will depend on the severity and course of thedisease, disorder or condition, previous therapy, the patient's healthstatus and response to the drugs, and the judgment of the treatingphysician. In one aspect, prophylactic treatments include administeringto a mammal, who previously experienced at least one symptom of thedisease being treated and is currently in remission, a pharmaceuticalcomposition comprising a compound disclosed herein, or apharmaceutically acceptable salt thereof, in order to prevent a returnof the symptoms of the disease or condition.

In certain embodiments wherein the patient's condition does not improve,upon the doctor's discretion the administration of the compounds areadministered chronically, that is, for an extended period of time,including throughout the duration of the patient's life in order toameliorate or otherwise control or limit the symptoms of the patient'sdisease or condition.

Once improvement of the patient's conditions has occurred, a maintenancedose is administered if necessary. Subsequently, in specificembodiments, the dosage or the frequency of administration, or both, isreduced, as a function of the symptoms, to a level at which the improveddisease, disorder or condition is retained. In certain embodiments,however, the patient requires intermittent treatment on a long-termbasis upon any recurrence of symptoms.

The amount of a given agent that corresponds to such an amount variesdepending upon factors such as the particular compound, diseasecondition and its severity, the identity (e.g., weight, sex) of thesubject or host in need of treatment, but nevertheless is determinedaccording to the particular circumstances surrounding the case,including, e.g., the specific agent being administered, the route ofadministration, the condition being treated, and the subject or hostbeing treated.

In general, however, doses employed for adult human treatment aretypically in the range of 0.01 mg-2000 mg per day. In one embodiment,the desired dose is conveniently presented in a single dose or individed doses administered simultaneously or at appropriate intervals,for example as two, three, four or more sub-doses per day.

In one embodiment, the daily dosages appropriate for the compounddisclosed herein, or a pharmaceutically acceptable salt thereof,described herein are from about 0.01 to about 50 mg/kg per body weight.In some embodiments, the daily dosage or the amount of active in thedosage form are lower or higher than the ranges indicated herein, basedon a number of variables in regard to an individual treatment regime. Invarious embodiments, the daily and unit dosages are altered depending ona number of variables including, but not limited to, the activity of thecompound used, the disease or condition to be treated, the mode ofadministration, the requirements of the individual subject, the severityof the disease or condition being treated, and the judgment of thepractitioner.

Toxicity and therapeutic efficacy of such therapeutic regimens aredetermined by standard pharmaceutical procedures in cell cultures orexperimental animals, including, but not limited to, the determinationof the LD₅₀ and the ED₅₀. The dose ratio between the toxic andtherapeutic effects is the therapeutic index and it is expressed as theratio between LD₅₀ and ED₅₀. In certain embodiments, the data obtainedfrom cell culture assays and animal studies are used in formulating thetherapeutically effective daily dosage range and/or the therapeuticallyeffective unit dosage amount for use in mammals, including humans. Insome embodiments, the daily dosage amount of the compounds describedherein lies within a range of circulating concentrations that includethe ED₅₀ with minimal toxicity. In certain embodiments, the daily dosagerange and/or the unit dosage amount varies within this range dependingupon the dosage form employed and the route of administration utilized.

In any of the aforementioned aspects are further embodiments in whichthe effective amount of the compound disclosed herein, or apharmaceutically acceptable salt thereof, is: (a) systemicallyadministered to the mammal; and/or (b) administered orally to themammal; and/or (c) intravenously administered to the mammal; and/or (d)administered by injection to the mammal; and/or (e) administeredtopically to the mammal; and/or (f) administered non-systemically orlocally to the mammal.

In any of the aforementioned aspects are further embodiments comprisingsingle administrations of the effective amount of the compound,including further embodiments in which (i) the compound is administeredonce a day; or (ii) the compound is administered to the mammal multipletimes over the span of one day.

In any of the aforementioned aspects are further embodiments comprisingmultiple administrations of the effective amount of the compound,including further embodiments in which (i) the compound is administeredcontinuously or intermittently: as in a single dose; (ii) the timebetween multiple administrations is every 6 hours; (iii) the compound isadministered to the mammal every 8 hours; (iv) the compound isadministered to the mammal every 12 hours; (v) the compound isadministered to the mammal every 24 hours. In further or alternativeembodiments, the method comprises a drug holiday, wherein theadministration of the compound is temporarily suspended or the dose ofthe compound being administered is temporarily reduced; at the end ofthe drug holiday, dosing of the compound is resumed. In one embodiment,the length of the drug holiday varies from 2 days to 1 year.

Combination Treatments

In certain instances, it is appropriate to administer at least onecompound disclosed herein, or a pharmaceutically acceptable saltthereof, in combination with one or more other therapeutic agents.

In one embodiment, the therapeutic effectiveness of one of the compoundsdescribed herein is enhanced by administration of an adjuvant (i.e., byitself the adjuvant has minimal therapeutic benefit, but in combinationwith another therapeutic agent, the overall therapeutic benefit to thepatient is enhanced). Or, in some embodiments, the benefit experiencedby a patient is increased by administering one of the compoundsdescribed herein with another agent (which also includes a therapeuticregimen) that also has therapeutic benefit.

In one specific embodiment, a compound disclosed herein, or apharmaceutically acceptable salt thereof, is co-administered with asecond therapeutic agent, wherein the compound disclosed herein, or apharmaceutically acceptable salt thereof, and the second therapeuticagent modulate different aspects of the disease, disorder or conditionbeing treated, thereby providing a greater overall benefit thanadministration of either therapeutic agent alone.

In any case, regardless of the disease, disorder or condition beingtreated, the overall benefit experienced by the patient is simply beadditive of the two therapeutic agents or the patient experiences asynergistic benefit.

For combination therapies described herein, dosages of theco-administered compounds vary depending on the type of co-drugemployed, on the specific drug employed, on the disease or conditionbeing treated and so forth. In additional embodiments, whenco-administered with one or more other therapeutic agents, the compoundprovided herein is administered either simultaneously with the one ormore other therapeutic agents, or sequentially.

In combination therapies, the multiple therapeutic agents (one of whichis one of the compounds described herein) are administered in any orderor even simultaneously. If administration is simultaneous, the multipletherapeutic agents are, by way of example only, provided in a single,unified form, or in multiple forms (e.g., as a single pill or as twoseparate pills).

The compounds disclosed herein, or a pharmaceutically acceptable saltthereof, as well as combination therapies, are administered before,during or after the occurrence of a disease or condition, and the timingof administering the composition containing a compound varies. Thus, inone embodiment, the compounds described herein are used as aprophylactic and are administered continuously to subjects with apropensity to develop conditions or diseases in order to prevent theoccurrence of the disease or condition. In another embodiment, thecompounds and compositions are administered to a subject during or assoon as possible after the onset of the symptoms. In specificembodiments, a compound described herein is administered as soon as ispracticable after the onset of a disease or condition is detected orsuspected, and for a length of time necessary for the treatment of thedisease. In some embodiments, the length required for treatment varies,and the treatment length is adjusted to suit the specific needs of eachsubject.

EXAMPLES Abbreviations

-   BINAP: (2,2′-bis(diphenylphosphino)-1,1′-binaphthyl);-   DCE: 1,2-dichloroethane;-   DCM: dichloromethane;-   DI: deionized;-   DIEA or DIPEA: diisopropylethylamine;-   EtOAc: ethyl acetate;-   EtOH: ethanol;-   equiv: equivalents, typically molar equivalents;-   HATU:    1-[bis(dimethylamino)methylene]-1H-1,2,3-triazolo[4,5-b]pyridinium    3-oxid hexafluorophosphate;-   IPA: isopropylalcohol;-   IPAc: isopropyl acetate;-   MeOAc: methyl acetate;-   NBS: N-bromosuccinimide;-   NCS: N-chlorosuccinimide;-   OPA: orthophosphoric acid;-   PTS: p-toluene sulfonic acid;-   Pd(Amphos)Cl₂:    bis(di-tert-butyl(4-dimethylaminophenyl)phosphine)dichloropalladium(II);-   Pd₂dba₃: tris(dibenzylideneacetone)dipalladium(0);-   Pd(dppf)Cl₂:    [1,1′-bis(diphenylphosphino)ferrocene]dichloropalladium(II);-   (pinB)₂: bis(pinacolato)diboron;-   rt or RT: room temperature;-   Rt: retention time;-   SFC: supercritical fluid chromatography;-   SST: somatostatin;-   SSTR: somatostatin receptor;-   TEA: trimethylamine;-   TFA: trifluoroacetic acid;-   THF: tetrahydrofuran;-   vol: volume, typically used for reaction volume or ratio of    solvents;-   Xphos: 2-dicyclohexylphosphino-2′,4′,6′-triisopropylbiphenyl;-   hrs: hours;-   h or hr: hour.

The following examples are provided for illustrative purposes only andnot to limit the scope of the claims provided herein.

Example 1: Synthesis of 3-Bromo-2-(methoxymethoxy)benzonitrile (Compound2b)

Diisoproylethylamine (114 mL, 1.3 equiv) was slowly added to a solutionof nitrile Compound 2 (100 g, 1 equiv) in CH₂Cl₂ (1 L, 10 vol) at 0° C.and stirred for 30 min. Chloromethyl methyl ether (MOMCl) (46 mL, 1.2equiv) was then added slowly while maintaining the internal temperatureat 0 to 5° C. The reaction was then allowed to warm to RT and stirredfor 4 h, until TLC showed complete reaction. The reaction was cooled to0° C. and quenched with DI-water (300 mL, 3 vol) and the layers wereseparated. The aqueous layer was extracted with CH₂Cl₂ (300 mL, 3 vol)and the combined organic layer was washed with water and brine,concentrated on a rotavap to afford 115 g of crude product as a brownoil. The crude product was purified on a plug of SiO2 and eluted with10% ethyl acetate and pet-ether (20 vol). Only one fraction wascollected, evaporated under vacuum, and dried under high vacuum toafford 86 g (70%) of Compound 2b as colorless oil, which was 99.96% pure(HPLC-AUC).

Example 2: Synthesis of (3-cyano-2-hydroxyphenyl)boronic Acid (Compound1a)

A solution of ^(i)PrMgCl in THF (2 M in THF, 340 mL, 2.2 equiv) wasadded slowly to a solution of Compound 2b (75 g, 1 equiv) in THF (1.12L, 15 vol) while maintaining the internal temperature at −5 to 5° C. andstirred for 10 min. Triisopropyl borate (180 mL, 2.5 equiv) was thenadded while maintaining the internal temperature at −5 to 3° C. Thereaction was then allowed to warm to room temperature and stirred for 18h until TLC showed complete reaction. The reaction was then cooled to−10° C. and quenched by slow addition of 3N HCl (620 mL, 6 equiv) at−10° C. The mixture was stirred for 3 h at RT and extracted with ethylacetate (525 ml, 5 vol) and the aqueous layer was extracted with ethylacetate (225 mL, 3 vol). The combined organic layer was successivelywashed with DI-water (3×3 vol), brine (525 mL, 3 vol) and concentratedunder vacuum to afford the crude material as a gummy solid. The crudewas stirred in pet-ether (525 mL, 5 vol) for 30 min and resulting solidswere filtered, washed with pet-ether (150 mL, 2 vol) and dried undervacuum to afford 35 g (70%) of boronic acid Compound 1a as an off whitesolid, which was 97.91% pure (HPLC-AUC).

Example 3: Synthesis of 2-Bromo-6-cyanophenyl acetate (Compound 2c)

3-Bromo-2-fluro-benzonitrile (25 g, 1.0 equiv), potassium acetate (5equiv) was mixed in DMSO (7 vol) and heated to 90-95° C. for 48 h. IPCshowed 0.38% of 3-bromo-2-fluro-benzonitrile and 96.5% of Compound 2.Reaction mixture was cooled to 25-30° C. and quenched with purifiedwater (25 vol water). Then pH was adjusted pH to 3-4 using 6N HClsolution. The obtained resulting mixture was diluted with MTBE (10 vol).The organic layer was separated, and aqueous layer was extracted with 10vol MTBE. Combined organic layers were washed with water (10 vol×3) andconcentrated to 2 vol level, chased with DCM (3 vol) and concentrateddown to 2 vol again before diluted with DCM (8 vol) to afford crudesolution of Compound 2. This solution was used in the subsequent processwithout further purification.

Crude solution of Compound 2 was mixed with acetic anhydride (1.3equiv), DMAP (0.1 equiv) at 25-30 C for 2 h with stirring. IPC showed0.8% Compound 2 and 94.8% Compound 2c. Reaction mass was diluted withpurified water (10 vol), stirred for 30 mins. Organic layer wasseparated. Aqueous layer was extracted with 2 vol DCM. The combinedorganic layer was washed with water (8 vol×2). Charcoal (10%) was addedto organic layer and stirred for 1 h before filtered through celite bed.The filtrate was then concentrated to 2 vol level, chased with 3 vol and2 vol of n-heptane subsequently before cooled to RT and stirred for 1 hrat 5-10° C. The product was isolated via filtration as solid (25.5 g,85% yield over two steps, HPLC purity 96.8%).

Example 4: Alternative Synthesis of 2-hydroxy-3-cyano-potassiumphenyltrifluoro borate (Compound 1c)

Compound 2c (20 g, 1.0 equiv) was mixed with KOAc (3.0 equiv),bis(pinacolato)diboron (1.2 equiv) in 2-methyl-THF. The mixture wasstirred and degassed with N₂ bubbling before addition of Pd(dppf)Cl₂.DCM(0.025 equiv). The resulting mixture was heated to 80-85° C. for 16 h.IPC showed 0.3% of starting material and 92% Compound 1b. Reactionmixture was cooled to 25-30° C. and filtered through celite pad. Thecelite pad was washed with MTBE (5 vol). The combined filtrate wasconcentrated to 2 vol and chased with MTBE to 2 vol. The resultingsolution was diluted with MTBE (10 vol) and stirred for 1 h at ambienttemperature. The suspension was again filtered through celite pad andthe celite pad was rinsed with MTBE. The combined filtrate was washedwith water (500 ml, 5 vol). The aqueous layer was extracted with MTBE.Combined organic layers were washed with 5% N-Acetyl-L-cysteine solutiontwice (each time 300 ml, 3 vol) and water (300 ml, 3 vol). Then theorganic layer was separated, treated with 10% active charcoal, filteredthrough celite pad. The celite pad was rinsed with MTBE. The combinedfiltrate was concentrated to 2 vol level, chased with methanol twice(2×4 vol) to 3 vol, and cooled to ambient temperature before used in thenext step.

Compound 1b solution was added with KHF₂ (5.0 equiv), purified water(2.6 vol), and MeOH (1 vol) before heated to 65° C. for 1 h. Thereaction mixture was diluted with MTBE (15 vol) before cooled to 10±5OC. The resulting suspension was stirred for 1 h before filtration. Thesolid was transferred to reaction flask, added with 20 vol acetone,stirred at 25±5° C. for 1 h, treated with 10% charcoal, and stirred foranother 1 h. The resulting reaction mixture was then filtered throughcelite pad. Filtrate was concentrated to 2 vol level, chased with MTBE(3 vol×2), concentrate to 2 vol level, and diluted with MTBE (4 vol).The suspension was stirred for 1 h at 25±5° C. before filtered to affordthe desired product Compound 1c as off-white solid (11.5 g, 49.5%, HPLCpurity 97.8%). ¹H NMR (400 MHz, DMSO-d₆): δ 8.2 (s, 1H), 7.41 (m, 1H),7.34 (m, 1H), 6.80 (m, 1H); ¹³C NMR (100 MHz, DMSO-d₆): 161.8, 138.0,138.0, 130.8, 119.4, 118.0, 96.6.

Example 5: Synthesis of 6-Bromo-3-chloroquinolin-4-ol (A-II)

N-Chlorosuccinimide (377 g, 1.05 equiv) was added to a suspension of6-bromoquinolin-4(1H)-one (Compound A-I, 600 g, 1 equiv) in acetic acid(12 L, 20 vol) at RT. The reaction was then heated to 50° C. and stirredfor 8 h. The reaction was cooled to 20° C., filtered, successivelywashed with AcOH (1.8 L, 3 vol), water (2.4 L, 4 vol), and MTBE (1.2 L,2 vol), and dried under vacuum on a filter to afford crude CompoundA-II. The crude material was stirred in MTBE (7.2 L, 12 vol) for 2 h,filtered, washed with MTBE (0.6 L, 1 vol) and dried under vacuum toafford 541 g (78%) of Compound A-II as an off white solid, which wasdetermined to be 97.35% pure (HPLC-AUC).

Example 6: Synthesis of 4,6-Dibromo-3-chloroquinoline (A-III)

Phosphorus tribromide (317 mL, 1.6 equiv) was slowly added to a solutionof A-II (540 g, 1 equiv) in DMF (7 L, 13 vol) at 0-5° C. The reactionwas allowed to warm to RT and stirred for 4 h. The reaction was cooledto 0° C. and quenched by sat. aqueous solution of NaHCO₃ to pH˜8 (10.8L, 20 vol) and diluted with water (5.4 L, 10 vol). The mixture wasstirred for 2 h at RT and the solids were filtered, washed with water(2.7 L, 5 vol), and dried on the filter under vacuum. The wet cake wasslurried in water (5.4 L, 10 vol) for 2 h and filtered, washed withwater (980 mL, 2 vol) and dried on the filter under vacuum to affordcrude Compound A-III as a solid. The crude material was stirred in MTBE(2.7 L, 5 vol) for 2 h, filtered, washed with MTBE (980 mL, 2 vol) anddried under vacuum to afford 434 g (65%) of Compound A-III as an offwhite solid, which was determined to be 97.95% pure (HPLC-AUC).

Example 7: Synthesis of tert-Butyl(1-(6-bromo-3-chloroquinolin-4-yl)piperidin-4-yl)carbamate (A-IV)

Diisopropylethylamine (932 mL, 4 equiv) and 4-(N-Boc-amino)piperidine(430 g, 1.6 equiv) were successively added to a solution of CompoundA-III (430 g, 1 equiv) in DMSO (4.3 L, 10 vol) at RT. The suspension wasthen heated to 140° C. and stirred for 3 h. The reaction was allowed tocool to RT, diluted with water (12.9 L, 30 vol), and stirred for 2 h.The resulting solids were filtered and dried on a filter. The wet cakewas dissolved in DCM (3 L, 7 vol), and the aqueous layer was extractedwith DCM (860 mL, 2 vol). The combined organic layer was washed withwater (2×2.1 L, 5 vol each), brine (2.1 L, 5 vol), and dried undervacuum to afford crude Compound A-IV as a solid. The crude material wasstirred in MTBE (2.21 L, 5 vol) for 1 h, filtered, washed with MTBE (860mL, 2 vol), and dried under vacuum to afford 412 g (70%) of CompoundA-IV as an off-white solid, which was determined to be 98.26% pure(HPLC-AUC).

Example 8: Alternative Synthesis of tert-Butyl(1-(6-bromo-3-chloroquinolin-4-yl)piperidin-4-yl)carbamate (A-IV)

A mixture of 6-bromo-4-chloro-quinoline (25 g, 1.0 equiv), DMF (6.0vol), 4-(tert-butoxy carbonylamino)piperidine (2.0 equiv), K₂CO₃ (2.5equiv), was stirred and heated to 105° C. for 16 h. Reaction wasmonitored by IPC-HPLC and showed 93.5% of Compound 4 and 0.12% ofCompound 3. Then reaction mixture was cooled to 25-30° C., added withpurified water (30 vol), and stirred for 2 h. Solid was filtered andwashed with purified water. The crude solid was slurred with n-heptane(5 vol), filtered, and washed with n-heptane (2 vol). The solid wasdried at 55° C. provided desired product Compound 4 (35.3 g, 84% yield).¹H NMR (400 MHz, DMSO-d₆): δ 8.69 (d, 1H), 8.02 (d, 1H), 7.88 (m, 1H),7.80 (m, 1H), 7.02 (d, 1H), 6.97 (d, 1H), 3.44 (m, 3H), 2.87 (m, 2H),1.94 (d, 2H), 1.68 (m, 2H), 1.38 (s, 9H).

Compound 4 (25 g, 1 equiv), DIPA (0.078 equiv), NCS (1.5 equiv), Toluene(10 vol) were mixed and heated at 70° C. for 4 h. Reaction mixture wasconcentrated to 3 vol level at 45±5° C., cooled to rt, diluted with MTBE(10 vol), washed with purified water (10 vol). After layer separation,aqueous layer was extracted with MTBE (5.0 vol). Combined organic layerswere washed with purified water twice (2×5 vol) followed by brine. Theorganic layer was dried over sodium sulfate, concentrated to 2 vollevel, chase with MTBE twice (2×2 vol). Then it was cooled and addedwith MTBE (1 vol) before warmed to 50±5° C. and stirred for 1 hour. Theresulting suspension was cooled to 5±5° C. and stirred for 1 h. Solidwas collected by filtration and washed with pre-cooled MTBE (1 vol).Above solid was taken in MTBE (2 vol) and heated to 55±5° C. again,stirred for 1 h, cooled to 5±5° C., and stirred for another 1 h. Solidwas collected by filtration and washed with pre-cooled MTBE (1 vol). Thecollected solid was dried under reduced pressure at 45±5° C. for 8 h toprovide product Compound A-IV with 98.9% HPLC purity and 66% (18 g)isolated yield. ¹H NMR (400 MHz, DMSO-d₆): δ 8.65 (s, 1H), 8.25 (d, 1H),7.92 (m, 1H), 7.73 (m, 1H), 3.76 (s, 1H), 3.51 (m, 2H), 3.37 (d, 2H),2.14 (d, 2H), 1.69 (m, 2H), 1.46 (s, 9H).

Example 9: Synthesis of tert-Butyl(1-(6-(3-cyano-2-hydroxyphenyl)-3-(3,5-difluorophenyl)quinolin-4-yl)piperidin-4-yl)carbamate(A-VI)

Quinoline A-IV (350 g, 1 equiv), (3-cyano-2-hydroxyphenyl)boronic Acid(Compound 1a) (155 g, 1.2 equiv), and K₂CO₃ (438 g, 4 equiv) werecharged to the round bottomed flask. 1,4-Dioxane (3.5 L, 10 vol) andDI-water (350 mL, 1 vol) were added to the flask and the resultingreaction mixture was degassed with argon for 30 min. PdCl₂(dppf).CH₂Cl₂(32.5 g, 0.05 equiv) was added to the reaction under argon and themixture was degassed further for 10 min. The reaction was stirred at80-85° C. and monitored by TLC and HPLC. After complete reaction (6 h),it was allowed to cool to 25-30° C. and 3,5-difluorophenylboronic acid(346 g, 3 equiv) was added to the reaction mixture which was thendegassed with argon for 10 min. PdCl₂(amphos) (25.9 g, 0.05 equiv) wasadded to the flask under argon atmosphere and the reaction mixture wasdegassed further for 10 min. The reaction was then heated to 90-100° C.and stirred for 19 h (monitored by TLC and HPLC). HPLC showed 82.04% ofCompound A-VI along with 1.95% of un-reacted Compound A-V and 0.94% ofanother impurity at 8.2 min. The reaction was allowed to cool to 25-30°C. and filtered through a pad of Celite and washed with ethyl acetate(1350 mL, 3 vol). The filtrate was concentrated under vacuum until ˜10%solvent remained and the resulting residue was diluted with ethylacetate (6.3 L, 18 vol), washed with water (2×3.5 L, 10 vol each), brine(3.5 L, 10 vol), and dried over anhydrous Na₂SO₄. The organic layer wasconcentrated under vacuum to dryness and was then slurried in ethylacetate (2.1 L, 6 vol) for 4 h at RT (after 2.5 h of stirring at RT,free solid formation was observed). Resulting free solids were filtered,washed with ethyl acetate (700 mL, 2 vol) and dried under vacuum untilconstant weight to afford 200 g (45%) of Compound A-VI as an off whitesolid, which was 98.4% pure (HPLC-AUC) with approx. 3500 ppm of tracePalladium.

Example 10: Alternative Synthesis of tert-Butyl(1-(6-(3-cyano-2-hydroxyphenyl)-3-(3,5-difluorophenyl)quinolin-4-yl)piperidin-4-yl)carbamate(A-VI)

Compound A-IV (25.0 g, 1 equiv), Compound 1c (1.2 equiv), K₂CO₃ (3.0equiv), 1,4-dioxane (9 vol), and purified water (0.75 vol) was added tothe reaction flask. The mixture was degassed with N₂ bubbling beforeaddition of Pd(PPh₃)₄ (0.017 equiv). Then the reaction mixture washeated to 80-85° C. for 12 h. IPC at 12 h showed <1% of Compound A-IV.Then 3,5 diflurophenylboronic acid (2.0 equiv), Pd(amphos)Cl₂ (0.03equiv) was added and the reaction mixture was degassed again beforeheated up to at 90-95° C. for 6 h. IPC showed <2% of remaining CompoundA-V. Pure Compound A-V sample was isolated and characterized by ¹H NMR(400 MHz, DMSO-d₆): δ 8.63 (s, 1H), 8.27 (bs, 1H), 7.94 (m, 2H), 7.53(d, 1H, J=7.2 Hz), 7.47 (d, 1H, J=6.0 Hz), 6.99 (d, 1H, J=7.6 Hz), 6.77(bs, 1H), 3.50 (m, 1H), 3.41 (m, 2H), 3.34 (m, 2H), 1.87 (m, 2H), 1.65(m, 2H), 1.39 (s, 9H).

Workup and Pd Removal:

The reaction mixture was cooled to 25-30° C., filtered through celitepad. The celite pad was washed with IPAc (2.0 vol). The filtrate wascombined, concentrated to 3 vol, chased with IPAc (5 vol) twice to 4vol. The resulting solution was diluted with IPAc (8 vol) and washedwith water (2×10 vol). Organic layer was separated and washed with 1%N-Acetyl L-cysteine (2×10 vol) before concentrated to 6 vol. Theresulting suspension was stirred at reflux for 2 h and cooled to ambienttemperature. The suspension is further cooled to 10±5° C., stirred for 2h, and filtered. The filter cake was washed with 1 vol IPAc and dried toprovide the desired crude product as pale-yellow solid.

Isolated Compound A-VI crude solid was dissolved in 2-methylTHF (15vol), added with SILIAMET S THIOL (0.25% w/w), and stirred at ambienttemperature for 3 h. The suspension was filtered through celite bed,washed with 2-methylTHF (2 vol). The above process was repeated again.The final filtrate was concentrated to 2 vol, chased with n-heptanetwice (2×3 vol). The resultant suspension was filtered. The solid wasdried under vacuum at 45±5° C. to afford the purified Compound A-VI assolid (HPLC purity 96% with Pd level of 13 ppm).

Example 11: Additional Purification of A-VI

Compound A-VI (200 g, 98.40% pure) was taken in IPAc (1 L, 5 vol) andrefluxed for 1 h. The mixture was then allowed to cool to RT and thencooled to 15° C., filtered, washed with IPAc (600 mL, 3 vol), and driedto afford 170 g of Compound A-VI as an off white solid, which was 98.71%pure (HPLC-AUC) with approx. 50 ppm of trace Palladium.

Example 12: Removal of Residual Palladium from A-VI

Compound A-VI (150 g, 98.71% pure) was dissolved in THF (3.4 L, 20 vol).Si-Thiol (240 g) was added and the solution stirred overnight at RT. Themixture was filtered through a Celite bed, washed with THF (510 mL, 3vol), and concentrated under vacuum to afford a solid. The crude solidwas then diluted with IPAc (1 L, 5 vol) and the slurry was refluxed for2 h. The mixture was then allowed to cool to RT, then was cooled to 15°C., filtered, washed with IPAc (510 mL, 3 vol), and dried to afford 150g of Compound A-VI as an off-white solid, which was 100% pure (HPLC-AUC)with no detectable residual Palladium.

Example 13:3-(4-(4-Amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl)-2-hydroxy-benzonitrile,di-HCl Salt (Compound A-2HCl)

A suspension of Compound A-VI (150 g, 269.5 mmol) in 6M HCl-IPA (1.45 L,10 vol) was stirred at RT for 4 h. TLC showed complete reaction (thicksuspension observed). The reaction was diluted with MTBE (2.17 L, 15vol), stirred for 2 h, and the resulting solids were filtered, washedwith MTBE (290 mL, 2 vol) and dried under vacuum to afford 130 g (91%)of Compound A-2HCl salt as a pale yellow solid, which was 100% pure(HPLC-AUC), with no detectable residual palladium. Chloride titrationshowed 15.8 wt % of chloride.

Example 14:3-(4-(4-Amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl)-2-hydroxy-benzonitrile,HCl Salt (Compound A-HCl)

A suspension of Compound A-VI (120.0 g) in IPAc (5 vol) was added with6M HCl in isopropyl acetate (10 vol) at rt. The resulting mixture wasstirred for 2 h at rt. After completion of the reaction (Compound A-VIcontent was found to be <1.0% by HPLC), the thick suspension was dilutedwith IPAc (0.6 L, 5 vol), stirred for 1 h and filtered. The filter cakewas washed with IPAc (240.0 mL, 2 vol) and suck dried under vacuum for 2h to afford Compound A-2HCl as a yellow solid. Crude Yield: 110.0 g(96%)

The solution of Compound A-2HCl (110.0 g) in purified water (18 vol) washeated to 45±5° C. before aq. NH₃ (25%, 2.4 equiv) was slowly added andstirred for 1 h at 45±5° C. The pH of the final reaction mixture was5-5.5. Then, purified water (550.0 mL, 5 vol) was added slowly at sametemperature over 2 h. The resulting suspension was cooled to rt then to10±5° C. and stirred for 1 h. The mixture was filtered and washed withcold purified water (2 vol) (10±5° C.). The wet cake was suck dried foran hour and dried at 55±5° C. for 24 h under vacuum to afford CompoundA-HCl as a bright yellow to gold color solid (88.0 g, 83% isolated yieldwith 99.4% HPLC purity).

Example 15: Preparation of Compound A-HCl from Compound A-2HCl

A suspension of Compound A-2HCl (118 g) in DI-water (2.4 L, 20 vol) wasstirred at 40-45° C. Aqueous ammonia (25%, 35.4 m, 0.3 mL/g) was slowlyadded to the hot suspension to adjust pH˜5-6 and stirred for 1 h.DI-water (590 mL, 5 vol) was added at same temperature over 2 h and thesuspension was allowed to cool to 25° C., then was further cooled to5-10° C. using ice-water. The free solids were then filtered and washedwith DI-water (3×500 mL). The wet cake was dried in a vacuum oven at 55°C. for 18 h to afford constant weight of 101 g (92%) of Compound A-HClas a pale yellow powder, which was 100% pure (HPLC-AUC), with nodetectable residual palladium. Chloride titration showed 8.1 wt % ofchloride.

Example 16: Alternate Preparation of Compound A-HCl from Compound A-2HCl

A suspension of Compound A-2HCl (2.1 g) in 1:1 IPA:water (10 mL, 5 vol)was stirred at 45° C. Concentrated ammonium hydroxide (28-30%, 0.8equiv) was slowly added and the hot suspension was stirred for 1 h.DI-water (20 mL, 10 vol) was added at same temperature over 2 h and thesuspension was cooled to 10° C. over 2 h. The resulting flowable slurrywas then filtered and washed with 15:85 IPA:water (2×3 vol). The wetcake was dried under vacuum at 45° C. for 18 h to afford constant weightof 1.42 g (78%) of Compound A-HCl as a pale yellow powder. Chloridetitration showed 8.0 wt % of chloride.

Example 17: Alternate Preparation of Compound A-HCl from CompoundA-2HCl, without the Use of a Base

About 80 mg of Compound A-2HCl salt was dissolved in various solvents(see below table). Reactions were heated to 50° C. for 30 minutes,cooled over 2 h to RT, held at RT for 1 h, then the solids were filteredand dried under vacuum at 50° C. Solvent was evaporated to yieldCompound A-HCl and the resulting XRPD pattern was evaluated. Eachsolvent system, except 9:1 methyl acetate:water, resulted in the XRPDpattern as in FIG. 1.

diHCl salt, mg Solvent(s) vol XRPD pattern 79.3 IPA:water (8:2) 10 FIG.1 76.7 IPA:water (7:3) 10 FIG. 1 78.3 methyl acetate:water (9:1) 10di-HCl 79.4 methyl acetate:water (8:2) 10 FIG. 1 78.7 water 6 FIG. 179.5 water 10 FIG. 1 92.7 water + 0.5 equiv NaHCO₃ 10 FIG. 1

Example 18: Preparation of Compound A from Compound A-2HCl

About 1.2 g of Compound A-2HCl was slurried in 15 volumes of sat. sodiumbicarbonate for 30 minutes. The solid was filtered and dried at 40-45°C. under vacuum to yield 730 mg (77%) of Compound A as the free base.

Example 19: Preparation of Compound A-HCl from Compound A

About 17 mg of Compound A, free base was dissolved in various solvents(500 uL each, see below table) followed by addition of 1.0 equiv of 3MHCl/IPA solution. The solids were filtered to yield Compound A-HCl andthe resulting XRPD pattern was evaluated. Each solvent, except MTBE,resulted in the XRPD pattern as in FIG. 1.

Free Base, mg HCl/IPA, equiv Solvent XRPD pattern 18.5 1 MeOAc FIG. 117.9 1 Acetonitrile FIG. 1 17.9 1 THF FIG. 1 16.1 1 EtOH:water (9:1)FIG. 1 17.1 1 Acetone:water (9:1) FIG. 1 17.2 1 MTBE amorphous

Example 20. X-Ray Powder Diffraction (XRPD)

X-ray Powder diffraction was done using a Rigaku MiniFlex 600. Sampleswere prepared on Si zero-return wafers. A typical scan is from 20 of 4to 30 degrees, with step size 0.05 degrees over five minutes with 40 kVand 15 mA. Typical parameters for XRPD are listed below.

Parameters for Reflection Mode X-ray wavelength Cu Kα1, 1.540598 Å,X-ray tube setting 40 kV, 15 mA Slit condition Variable + Fixed SlitSystem Scan mode Continuous Scan range (°2TH) 4-30 Step size (°2TH) 0.05Scan speed (°/min) 5Characterization of Crystalline Compound A-HCl

The X-Ray powder diffraction pattern for Compound A-HCl is displayed inFIG. 1. This XRPD pattern shows a crystalline form of Compound A-HCl.Peaks include the peaks listed in the following table:

Angle 2-Theta (°) Rel. Intensity (%) 4.5 50 9.1 50 10.2 100 16.3 52 18.457 19.1 55 20.7 42 23.3 43 23.4 42 23.6 47 27.1 40 28.0 34Characterization of the Crystalline Compound A-2HCl

The X-Ray powder diffraction pattern for Compound A-2HCl is displayed inFIG. 4. This XRPD pattern shows a crystalline form of Compound A-2HCl.Peaks include the peaks listed in the following table:

Angle 2-Theta (°) Rel. Intensity (%) 5.4 33 7.3 100Characterization of the Pattern A of Crystalline Compound A

The X-Ray powder diffraction pattern for Pattern A of Compound A isdisplayed in FIG. 7(a). This XRPD pattern shows a crystalline form ofCompound A. Peaks include the peaks listed in the following table:

Angle 2-Theta (°) Peak relative intensity, % 9.2 100 12.3 35 14.4 3124.0 29Characterization of the Pattern B of Crystalline Compound A

The X-Ray powder diffraction pattern for Pattern B of Compound A isdisplayed in FIG. 7(b). This XRPD pattern shows a crystalline form ofCompound A. Peaks include the peaks listed in the following table:

Angle 2-Theta (°) Peak relative intensity, % 5.9 34 13.9 22 14.2 20 17.5100 24.6 30Characterization of the Pattern C of Crystalline Compound A

The X-Ray powder diffraction pattern for Pattern C of Compound A isdisplayed in FIG. 7(c). This XRPD pattern shows a crystalline form ofCompound A. Peaks include the peaks listed in the following table:

Angle 2-Theta (°) Peak relative intensity, % 7.2 41 8.3 100 10.9 26 12.077

Example 21. Solubility in Water, FaSSGF, and FaSSIF

The salt forms were slurried in water, Fasted State Simulated GastricFluid (FaSSGF), and Fasted State Simulated Intestinal Fluid (FaSSIF) at37° C. for 2 days. Part of the slurry was filtered using a 0.4 uMsyringe filter to recover the supernatant for solubility measurement andthe rest was filtered to recover the remaining solid for XRPD analysis.

A calibration curve was developed for measuring concentration via HPLC.Compound A was added to a 100 mL volumetric flask and then filled to thevolume with HPLC grade ethanol. Table 3 shows the calibrationconcentration and corresponding AUC. A 30 minute gradient method withSolvent A as water with 0.1% TFA and solvent B as Methanol:Acetonitrile(1:1) was used.

TABLE 3 Concentration, mg/mL AUC (at 261 nM) 0.12 1096.75 0.087 787.020.034 427.95 0.0063 55.46 0.00013 1.99

The calibration correlation was used to measure the solubility of thesalt forms. From the clear saturated solution, 0.1 mL liquid wastransferred into a 10 mL vial and then filled to the volume withmethanol. The solution was then directly injected into the HPLC.

Results from the solubility studies are in Table 4.

TABLE 4 Water, Compound mg/mL FaSSGF, mg/mL FaSSIF, mg/mL Compound A(free base) 0.005 2.43 0.013 Compound A-HCl 0.83 2.12 0.01 CompoundA-2HCl 5.63 3.69 0.011

The solubility is clearly a function of pH. It was also found that thecompounds remained as an HCl salt in FaSSGF which simulates the stomachfluid.

Example 22: Dynamic Vapor Sorption (DVS)

Dynamic Vapor Sorption (DVS) was done using a DVS Intrinsic 1. Thesample is loaded into a sample pan and suspended from a microbalance. Atypical sample mass for DVS measurement is 25 mg. Nitrogen gas bubbledthrough distilled water provides the desired relative humidity.

A typical measurement includes the steps:

-   -   1—Equilibrate at 50% RH    -   2—50% to 2%. (50%, 40%, 30%, 20%, 10% and 2%)        -   a. Hold minimum of 5 mins and maximum of 60 minutes at each            humidity. The pass criteria is less than 0.002% change    -   3—2% to 95% (2%, 10%, 20%, 30%, 40%, 50%, 60%, 70%, 80%, 90%,        95%)        -   a. Hold minimum of 5 mins and maximum of 60 minutes at each            humidity. The pass criteria is less than 0.002% change    -   4—95% to 2% (95%, 80%, 70%, 60%, 50%, 40%, 30%, 20%, 10%, 2%)        -   a. Hold minimum of 5 mins and maximum of 60 minutes at each            humidity. The pass criteria is less than 0.002% change    -   5—2% to 50% (2%, 10%, 20%, 30%, 40%, 50%)        -   a. Hold minimum of 5 mins and maximum of 60 minutes at each            humidity. The pass criteria is less than 0.002% change

As noted below, Compound A-HCl picked up much less moisture thanCompound A-2HCl.

Compound A-HCl

Testing the mono-HCl salt of Compound A (Compound A-HCl) showed areversible water uptake (˜4.5% w/w) between 2 and 95% RH; and areversible water uptake (˜2.3% w/w) between 15 and 75% RH. The XRPD wasunchanged after DVS analysis (FIG. 9).

Compound A-2HCl

Testing the di-HCl salt of Compound A (Compound A-2HCl) showed areversible water uptake (˜18% w/w) between 2 and 95% RH; and areversible water uptake (˜9% w/w) between 15 and 75% RH. The XRPD wasunchanged after DVS analysis (FIG. 10).

Example 23: Thermal Study of Compound A-HCl

Differential Scanning Calorimetry (DSC) was done using a Mettler ToledoDSC3+. Aluminum crucible with cramped pinned lid were used. The heatingrates are usually 10° C. per minute from 30 to 250° C.

Thermogravimetric Analysis (TGA) was done using a Mettler ToledoTGA/DSC3+. Aluminum crucible with cramped pinned lid were used. Theheating rates are usually 10° C. per minute from 30 to 250° C.

Protective and purge gasses are nitrogen (20-30 mL/min and 50-100mL/min). Typical parameters for DSC/TGA are listed below.

Parameters Method Ramp Sample size 5-10 mg Heating rate 10.0° C./minTemperature range 30 to 250° C.Compound A-HCl

As shown in FIG. 2(a) and FIG. 2(b), Compound A-HCl shows multiple peaksin its thermogram. These peaks were evaluated using thermal analysis andXRPD techniques. Compound A-HCl was heated to various temperatures usingTGA/DSC followed by cooling to room temperature and analysis using XRPD.The solid was analyzed by Karl Fischer titration and showed 5.9% waterin the sample. The following thermal treatments were performed:

Sample 1: heated the salt to 100° C. and cooled to RT followed by XRPD;

Sample 2: heated the salt to 150° C. and cooled to RT followed by XRPD;

Sample 3: heated the salt to 200° C. and cooled to RT followed by XRPD;

Sample 4: heated the salt to 250° C. and cooled to RT followed by XRPD;and

Sample 5: heated the salt to 295° C. and cooled to RT followed by XRPD.

It was observed that the solid is in tact up to 200° C. and melts at220° C., at which point the compound loses its chemical and crystalintegrity. Therefore, peaks above 220° C. are not relevant.

Since the solid contains about 5.9% water, the initial endothermic peakis due to water loss.

A DSC thermogram for Compound A-2HCl is shown in FIG. 5(a).

A TGA thermogram for Compound A-2HCl is shown in FIG. 5(b).

A DSC thermogram for Pattern C Compound A, free base is shown in FIG.8(a).

A TGA thermogram for Pattern C Compound A, free base is shown in FIG.8(b).

Example 24: Infrared (IR) Spectroscopy

200 mg of freshly dried potassium bromide was weighed and transferredinto a mortar and was ground into a fine powder. To this, 2.0 mg of testcompound was added and the solids were mixed thoroughly. A smallquantity of the powder was formed into a thin semi-transparent pellet. AShimadzu IR Prestige 21 was used to acquire the IR spectra of the testcompounds with 60 scans from 4000 cm⁻¹ to 400 cm⁻¹. Air was used as areference.

Characterization of the Crystalline Form of Compound A-HCl

The IR spectrum for Compound A-HCl is displayed in FIG. 3.Characteristic peaks include peaks at: 2223 cm⁻¹, 1620 cm⁻¹, 1595 cm⁻¹,1457 cm⁻¹, 1238 cm⁻¹, 1220 cm⁻¹, and 1117 cm⁻¹.

Characterization of the Crystalline Form of Compound A-2HCl

The IR spectrum for Compound A-2HCl is displayed in FIG. 6.Characteristic peaks include peaks at: 2227 cm⁻¹, 1620 cm⁻¹, 1594 cm⁻¹,1456 cm⁻¹, 1439 cm⁻¹, 1321 cm⁻¹, and 1122 cm⁻¹.

Example 25: High-Performance Liquid Chromatography (HPLC) Methods

Method A.

A Hitachi HPLC equipped with DAD detector was used for solubility andpurity tests. The HPLC column used was C18 5μ 100A, 4.6 mm×250 mm.Chromatographic conditions were as in the following tables:

Mobile Phase (A) 0.1% TFA in 1 L Milli Q water Mobile Phase (B)Acetonitrile Sample Temperature 25° C. Flow Rate 0.8 mL/min DetectionWavelength 232 nm Injection Volume 5 uL Run Time 25 min Injection Delay5 min

Step Time (min) Mobile Phase A (%) Mobile Phase B (%) 1 0.1 98 2 2 16 298 3 19 2 98 4 22 98 2 5 25 98 2Method B.

A Waters Alliance HPLC (or equivalent) equipped with DAD detector wasused for purity tests. The HPLC column used was C18 5μ 110A, 4.6 mm×250mm. Chromatographic conditions were as in the following tables:

Mobile Phase (A) 0.1% OPA in 1 L Milli Q water Mobile Phase (B)Acetonitrile Sample Temperature 25° C. Flow Rate 1.2 mL/min DetectionWavelength 232 nm/212 nm Injection Volume 5 uL Run Time 25 min InjectionDelay 5 min

Step Time (min) Mobile Phase A (%) Mobile Phase B (%) 1 0.1 98 2 2 0.598 2 3 7 2 98 4 17 2 98 5 21 98 2 6 25 98 2

Example 26:2-{4-[(4aS,8aS)-Octahydro-1H-pyrido[3,4-b][1,4]oxazin-6-yl]-3-(3-fluorophenyl)quinolin-6-yl}-4-methylpyridin-3-amine(1-8)

Step 1-1. Preparation of benzyl(4aS,8aS)-6-[3-chloro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)quinolin-4-yl]-octahydro-1H-pyrido[3,4-b][1,4]oxazine-1-carboxylate

To a dioxane solution (3.0 mL) of benzyl(4aS,8aS)-6-(6-bromo-3-chloroquinolin-4-yl)-octahydro-1H-pyrido[3,4-b][1,4]oxazine-1-carboxylate(1.0 equiv, 0.29 mmol, 150 mg) was added4,4,5,5-tetramethyl-2-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)-1,3,2-dioxaborolane(2.0 equiv, 0.58 mmol, 147 mg), KOAc (3.0 equiv, 0.87 mmol, 85 mg), andPd(dppf)Cl₂ (0.06 equiv, 0.02 mmol, 12 mg) under atmospheric N₂. Theresulting mixture was heated at 90° C. for 1 h and then cooled down toambient temperature. The reaction solution was concentrated under vacuumto afford 170 mg of the crude product. This material was used for nextstep without further purification.

Step 1-2. Preparation of benzyl(4aS,8aS)-6-[6-(3-amino-4-methylpyridin-2-yl)-3-chloroquinolin-4-yl]-octahydro-1H-pyrido[3,4-b][1,4]oxazine-1-carboxylate

To a dioxane solution (4.0 mL) of crude benzyl(4aS,8aS)-6-[3-chloro-6-(4,4,5,5-tetramethyl-1,3,2-dioxaborolan-2-yl)quinolin-4-yl]-octahydro-1H-pyrido[3,4-b][1,4]oxazine-1-carboxylate(1.0 equiv, 0.29 mmol, 170 mg) was added 2-bromo-4-methylpyridin-3-amine(1.0 equiv, 0.29 mmol, 56 mg), Pd(Amphos)Cl₂ (0.10 equiv, 0.029 mmol, 21mg), K₂CO₃ (3.0 equiv, 0.90 mmol, 125 mg), and water (0.4 mL) underatmospheric N₂. The resulting mixture was heated at 90° C. for 2 h. Thereaction mixture was cooled down to ambient temperature and concentratedunder vacuum. The residue obtained was purified by silica gelchromatography eluting with ethyl acetate/petroleum ether (3:1) toafford 70 mg of the desired product as yellow oil. MS (M+H)⁺=544.2.

Step 1-3. Preparation of benzyl(4aS,8aS)-6-[6-(3-amino-4-methylpyridin-2-yl)-3-(3-fluorophenyl)quinolin-4-yl]-octahydro-1H-pyrido[3,4-b][1,4]oxazine-1-carboxylate

To a toluene solution (3.0 mL) of benzyl(4aS,8aS)-6-[6-(3-amino-4-methylpyridin-2-yl)-3-chloroquinolin-4-yl]-octahydro-1H-pyrido[3,4-b][1,4]oxazine-1-carboxylate(1.0 equiv, 0.13 mmol, 70 mg) was added (3-fluorophenyl)boronic acid(2.0 equiv, 0.26 mmol, 36 mg), Pd(Amphos)Cl₂ (0.10 equiv, 0.013 mmol, 9mg), K₂CO₃ (3.0 equiv, 0.38 mmol, 53 mg), and water (0.3 mL) underatmospheric N₂. The resulting mixture was heated at 110° C. for 1 h. Thereaction mixture was cooled down to ambient temperature and concentratedunder vacuum. The residue obtained was purified by silica gelchromatography eluting with ethyl acetate/petroleum ether (1:1) toafford 50 mg of the desired product as yellow oil. MS (M+H)⁺=604.3.

Step 1-4. Preparation of2-{4-[(4aS,8aS)-octahydro-1H-pyrido[3,4-b][1,4]oxazin-6-yl]-3-(3-fluorophenyl)quinolin-6-yl}-4-methylpyridin-3-amine

Benzyl(4aS,8aS)-6-[6-(3-amino-4-methylpyridin-2-yl)-3-(3-fluorophenyl)quinolin-4-yl]-octahydro-H-pyrido[3,4-b][1,4]oxazine-1-carboxylate(1.0 equiv, 0.08 mmol, 50 mg) was combined with trifluoroacetic acid(2.0 mL) and the resulting mixture was heated at 80° C. for 2 h. Thereaction solution was cooled down to ambient temperature andconcentrated under vacuum. The residue obtained was purified byPrep-HPLC with the following conditions (Prep-HPLC-007): Column, SunFirePrep C18 OBD Column, 19*150 mm, 5 um, 10 nm; mobile phase, Water (0.1%TFA) and ACN (2.0% ACN up to 20.0% in 6 min, hold at 95% for 1 min, downto 2.0% in 1 min, hold at 2.0% for 1 min); Detector, UV 220 nm. Thisresulted in 11.4 mg (27%) of formic acid salt of2-{4-[(4aS,8aS)-octahydro-1H-pyrido[3,4-b][1,4]oxazin-6-yl]-3-(3-fluorophenyl)quinolin-6-yl}-4-methylpyridin-3-amineas a yellow solid. MS (M+H)⁺=470.2.

The following compounds were prepared similarly to Example 25 withappropriate substituting of reagents and substrates at different stepsand additional functional group modifications via well-known chemistrywith appropriate reagents as required. Different salts such as HCl orformic acid may be obtained through conventional methods.

Compound no. MS (M + H)⁺ 1-1 517.2 1-2 527.3 1-3 547.1 1-4 495.2 1-5515.2 1-6 517.2 1-7 481.2 1-9 471.2 1-10 471.2

Example A-1: Parenteral Pharmaceutical Composition

To prepare a parenteral pharmaceutical composition suitable foradministration by injection (subcutaneous, intravenous), 1-100 mg of awater-soluble salt of a compound disclosed herein, or a pharmaceuticallyacceptable salt or solvate thereof, is dissolved in sterile water andthen mixed with 10 mL of 0.9% sterile saline. A suitable buffer isoptionally added as well as optional acid or base to adjust the pH. Themixture is incorporated into a dosage unit form suitable foradministration by injection

Example A-2: Oral Solution

To prepare a pharmaceutical composition for oral delivery, a sufficientamount of a compound disclosed herein, or a pharmaceutically acceptablesalt thereof, is added to water (with optional solubilizer(s), optionalbuffer(s) and taste masking excipients) to provide a 20 mg/mL solution.

Example A-3: Oral Tablet

A tablet is prepared by mixing 20-50% by weight of a compound disclosedherein, or a pharmaceutically acceptable salt thereof, 20-50% by weightof microcrystalline cellulose, 1-10% by weight of low-substitutedhydroxypropyl cellulose, and 1-10% by weight of magnesium stearate orother appropriate excipients. Tablets are prepared by directcompression. The total weight of the compressed tablets is maintained at100-500 mg.

Example A-4: Oral Capsule

To prepare a pharmaceutical composition for oral delivery, 10-500 mg ofa compound disclosed herein, or a pharmaceutically acceptable saltthereof, is optionally mixed with starch or other suitable powderblends. The mixture is incorporated into an oral dosage unit such as ahard gelatin capsule, which is suitable for oral administration.

In another embodiment, 10-500 mg of a compound disclosed herein, or apharmaceutically acceptable salt thereof, is placed into Size 4 capsule,or size 1 capsule (hypromellose or hard gelatin) and the capsule isclosed.

Example A-5: Topical Gel Composition

To prepare a pharmaceutical topical gel composition, a compounddisclosed herein, or a pharmaceutically acceptable salt thereof, ismixed with hydroxypropyl celluose, propylene glycol, isopropyl myristateand purified alcohol USP. The resulting gel mixture is then incorporatedinto containers, such as tubes, which are suitable for topicaladministration.

Example B-1: SSTR Assays

Functional Assay for SSTR2 Agonists

General overview: All five SSTR subtypes are Gi coupled G-proteincoupled receptors (GPCRs) that lead to decreases in intracellular cyclicAMP (cAMP) when activated by an agonist. Therefore, measurement ofintracellular cAMP levels can be used to assess whether compounds of theinvention are agonists of SSTR subtypes (John Kelly, Troy Stevens, W.Joseph Thompson, and Roland Seifert, Current Protocols in Pharmacology,2005, 2.2.1-2.2). Human SSTR2 intracellular cAMP assay is describedbelow. The human SSTR1, 3, 4 and 5 assays follow the same protocol ofSSTR2.

cAMP Assay Protocol:

Four days prior to the assay, 5,000 Chinese hamster ovary cells (CHO-K1,ATCC #CCL-61) stably expressing the human SSTR2 are plated in each wellof a 96-well tissue culture-treated plate in Ham's F12 growth media(ThermoFisher #10-080-CM) supplemented with 10% donor bovine serum(Gemini Bio-Products #100-506), 100 U/mL penicillin; 100 ug/mLstreptomycin; 2 mM L-glutamine (Gemini Bio-Products #400-110) and 0.2mg/mL hygromycin B (GoldBio #31282-04-9). The cells are cultured at 37°C., 5% CO₂ and 95% humidity. On the day of the assay, the media isaspirated and the cells are treated with 50 μL of 1.6 μM NKH477 (Sigma#N3290) plus various dilutions of compounds of the invention in assaybuffer [1× Hank's Balanced Salt Solution (ThermoFisher #SH3058802), 0.5mM HEPES pH 7.4, 0.1% bovine serum albumin, 0.2 mM3-Isobutyl-1-methylxanthine (IBMX, VWR #200002-790)]. The cells areincubated for 20 minutes at 37° C. (the final concentration of thecompounds of the invention are typically 0-10,000 nM). The cells aretreated with 50 μL of lysis buffer (HRTF cAMP kit, Cisbio). The lysateis transferred to 384-well plates and cAMP detection and visualizationantibodies are added and incubated for 1-24 hours at room temperature.The time-resolved fluorescent signal is read with a Tecan M1000Promultiplate reader. The intracellular cAMP concentrations are calculatedby regression to a standard curve and are plotted vs. the concentrationof the compounds of the invention and the EC₅₀ of the compounds arecalculated using standard methods. All data manipulations are inGraphPad Prism v6.

Illustrative biological activity of compounds is demonstrated in thefollowing table by evaluating the inhibition of cAMP activities viahuman SST2R, where A means <10 nM; B means ≥10 nM and <100 nM; C means≥100 nM and <1000 nM; D means ≥1000 nM

Compound no. EC₅₀ Compound A A 1-1 A 1-2 A 1-3 A 1-4 A 1-5 A 1-6 A 1-7 A1-8 A 1-9 A 1-10 A

The examples and embodiments described herein are for illustrativepurposes only and various modifications or changes suggested to personsskilled in the art are to be included within the spirit and purview ofthis application and scope of the appended claims.

What is claimed is:
 1. A method of treating acromegaly or aneuroendocrine tumor, or combination thereof, in a human comprisingorally administering to the human with acromegaly or a neuroendocrinetumor, or combination thereof, a pharmaceutical composition comprisingthe compound3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrilemonohydrochloride, or solvate thereof; and at least one pharmaceuticallyacceptable excipient, wherein3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrilemonohydrochloride, or solvate thereof is crystalline and ischaracterized as having: an X-ray powder diffraction (XRPD) pattern withpeaks at 4.5° 2-Theta, 9.1° 2-Theta, 10.2° 2-Theta, 16.3° 2-Theta, 18.4°2-Theta, and 19.1° 2-Theta; an X-ray powder diffraction (XRPD) patternsubstantially the same as shown in FIG. 1; a Differential Scanningcalorimetry (DSC) thermogram with an endotherm having an onset at about207° C. and a peak at about 220° C.; a Differential Scanning calorimetry(DSC) thermogram substantially the same as shown in FIG. 2(a); aninfrared (IR) spectrum with peaks at 2223 cm⁻¹, 1620 cm⁻¹, 1595 cm⁻¹,1457 cm⁻¹, 1238 cm⁻¹, 1220 cm⁻¹, and 1117 cm⁻¹; an infrared (IR)spectrum substantially the same as shown in FIG. 3; an unchanged XRPDwhen heated up to about 200° C., upon exposure to more than 90% relativehumidity for about 24 hours, or upon exposure to about 75% RH and 40° C.over one week, or combinations thereof; or combinations thereof.
 2. Themethod of claim 1, wherein the pharmaceutical composition comprises1-100 mg of3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrilemonohydrochloride, or solvate thereof.
 3. The method of claim 1, whereinthe pharmaceutical composition is orally administered to the human oncea day.
 4. The method of claim 1, wherein the pharmaceutical compositionis in the form of a solid form pharmaceutical composition.
 5. The methodof claim 1, wherein the pharmaceutical composition is in the form of acapsule.
 6. The method of claim 5, wherein the capsule is orallyadministered to the human once a day.
 7. The method of claim 1, whereinthe pharmaceutical composition is in the form of a capsule and comprises10 mg of3-[4-(4-amino-piperidin-1-yl)-3-(3,5-difluoro-phenyl)-quinolin-6-yl]-2-hydroxy-benzonitrilemonohydrochloride, or solvate thereof.
 8. The method of claim 7, whereinthe capsule is orally administered to the human once a day.
 9. Themethod of claim 1, wherein the pharmaceutical composition is in the formof a tablet.
 10. The method of claim 9, wherein the tablet is orallyadministered to the human once a day.